Inhibitors of α-amino-β-carboxymuconic acid semialdehyde decarboxylase

ABSTRACT

The present disclosure discloses compounds capable of modulating the activity of α-amino-β-carboxymuconic acid semialdehyde decarboxylase (ACMSD), which are useful for the prevention and/or the treatment of diseases and disorders associated with defects in NAD +  biosynthesis, e.g., metabolic disorders, neurodegenerative diseases, chronic inflammatory diseases, kidney diseases, and diseases associated with ageing. The present application also discloses pharmaceutical compositions comprising said compounds and the use of such compounds as a medicament.

RELATED APPLICATIONS

This application claims the benefit of and priority to U.S. provisionalapplication No. 62/043,853, filed Aug. 29, 2014, the entire contents ofwhich are incorporated herein by reference in its entirety.

FIELD OF THE DISCLOSURE

The present disclosure relates to compounds capable of modulating theactivity of α-amino-β-carboxymuconic acid semialdehyde decarboxylase(ACMSD). The compounds of the disclosure may be used in methods for theprevention and/or the treatment of diseases and disorders associatedwith defects in NAD⁺ biosynthesis, e.g., metabolic disorders,neurodegenerative diseases, chronic inflammatory diseases, kidneydiseases, and diseases associated with ageing.

BACKGROUND OF THE DISCLOSURE

ACMSD is a critical enzyme for tryptophan metabolism, and regulates NAD⁺biosynthesis from tryptophan. ACMSD is a zinc-dependent amidohydrolasethat participates in picolinic acid (PA), quinolinic acid (QA) and NADhomeostasis. ACMSD stands at a branch point of the NAD⁺ biosyntheticpathway from tryptophan and determines the final fate of the amino acid,i.e., transformation into PA, complete oxidation through the citric acidcycle, or conversion into NAD⁺ through QA synthesis.

ACMSD has been purified from liver, kidney, and brain human tissues.There are two isoforms ACMSD1 and ACMSD2 derived from a differentialsplicing of ACMSD gene transcription but only ACMSD1 is endowed withenzymatic activity. ACMSD1 directs ACMS (α-amino-ω-carboxymuconic acidsemialdehyde) to the acetyl-CoA pathway, and when ACMSD1 is inhibited,ACMS is non-enzymatically converted to quinolinic acid (QA) leading tothe formation of NAD⁺ and an increase in the intracellular level ofNAD⁺.

Increased levels of NAD⁺ have been shown to protect against neuronaldegeneration, improve muscle function and oxidative metabolism in mice,and enhance lifespan in worms. Whilst reduced levels of NAD⁺ have beenassociated with a range of pathophysiological states including type 2diabetes (T2D), hyperlipidemia (elevated cholesterol and TAGs),mitochondrial diseases, neutropenia, cancers, and kidney disorders.

The inhibition of ACMSD thus represents a novel approach to increaseNAD⁺ levels and modify disease pathophysiologies associated with defectsin NAD⁺ biosynthesis.

SUMMARY OF THE DISCLOSURE

It is an object of embodiments of the disclosure to provide novel seriesof compounds capable of modulating the activity ofα-amino-β-carboxymuconic acid semialdehyde decarboxylase (ACMSD), whichcompounds are useful for the prevention and/or the treatment of diseasesand disorders associated with defects in NAD⁺ biosynthesis, e.g.,metabolic disorders, neurodegenerative diseases, chronic inflammatorydiseases, kidney diseases, and diseases associated with ageing.

Compounds of Formula (I), as defined herein, may be used in thetreatment of a disease or disorder in which ACMSD plays a role. Thedisclosure features methods of treating a disease or disorder associatedwith ACMSD dysfunction or with abnormalities in NAD⁺ biosynthesis byadministering to subjects suffering from or susceptible to developing adisease or disorder associated with ACMSD dysfunction a therapeuticallyeffective amount of one or more compounds that increases intracellularNAD⁺ by ACMSD1 inhibition, in an amount sufficient to activate sirtuins(SIRTs) and the downstream targets of SIRTs, such as PGC-1α, FoxO1and/or superoxide dismutase (SOD). The methods of the present disclosurecan be used in the treatment of ACMSD dependent diseases by inhibitingACMSD. Inhibition of ACMSD may provide a novel approach to theprevention and treatment of metabolic disorders, neurodegenerativediseases, chronic inflammatory diseases, kidney diseases, diseasesassociated with ageing and other ACMSD dependent diseases, or diseasescharacterized by defective NAD⁺ synthesis.

Accordingly, a first aspect of the present disclosure relates to acompound represented by Formula (I):

or a pharmaceutically acceptable salt or tautomer thereof,

wherein:

X is O, OH, or Cl;

L is —(CH₂)_(m)CH₂CH₂—, —(CH₂)_(m)Y(CH₂)_(p)—, —(CH₂)_(m)C(O)(CH₂)_(p)—,—(CH₂)_(m)C(O)O(CH₂)_(p)—, —(CH₂)_(m)C(O)NR²(CH₂)_(p)—, or—(CH₂)_(m)NR²C(O)(CH₂)_(p);

Y is O, N or S(O)_(q);

R¹ is C₆-C₁₀ aryl or heteroaryl, wherein the aryl and heteroaryl aresubstituted with R^(a) and R^(b), and optionally substituted with one ormore R^(e);

R² is H or C₁-C₆ alkyl;

one of R^(a) and R^(b) is hydrogen and the other is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)tetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r) isoxazol-3-ol,—(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH, —(CH₂)_(r)C(O)NHCN, or—(CH₂)_(r)C(O)NHS(O)₂alkyl;

R^(c) is H, C₁-C₆ alkyl, C₁-C₆ haloalkyl, halogen, —CN, —OR^(x),—CO₂R^(x), or NO₂;

R^(d) is methyl, optionally substituted 5- to 10-membered aryl,optionally substituted 5- or 6-membered heteroaryl, or optionallysubstituted 5- or 6-membered carbocycle;

each R^(x) is independently at each occurrence hydrogen or C₁-C₆ alkyl;

each R^(e) is independently C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,halogen, —OR^(y), C₁-C₆ haloalkyl, —NHR^(z), —OH, or —CN;

R^(f) is H or absent;

each R^(y) and R^(z) is independently hydrogen, C₁-C₆ alkyl, or C₁-C₆haloalkyl;

each m and p independently is 0, 1 or 2, wherein m+p<3;

q is 0, 1, or 2;

r is 0 or 1; and

the dotted line is an optional double bond;

with the proviso that R^(c) is not hydrogen or —CN when X is O, L is—SCH₂— and R^(d) is optionally substituted phenyl, R^(c) is not C₁-C₆alkyl when X is O, L is —SCH₂— and R^(d) is methyl, and that R^(c) isnot —CN when X is O, L is —SCH₂— and R^(d) is 2-furyl.

A second aspect of the present disclosure relates to pharmaceuticalcompositions comprising a compound of Formula (I), or a pharmaceuticallyacceptable salt thereof, and at least one of a pharmaceuticallyacceptable carrier, diluent, or excipient.

A third aspect of the disclosure relates to a method of treating adisease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction comprising administering to the subject suffering from orsusceptible to developing a disease or disorder associated with ACMSDdysfunction a therapeutically effective amount of one or more compoundsof Formula (I).

A fourth aspect of the disclosure relates to a method of preventing adisease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction comprising administering to the subject suffering from orsusceptible to developing a disease or disorder associated with ACMSDdysfunction a therapeutically effective amount of one or more compoundsof Formula (I).

A fifth aspect of the disclosure relates to a method of reducing therisk of a disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction comprising administering to the subject suffering from orsusceptible to developing a disease or disorder associated with ACMSDdysfunction a therapeutically effective amount of one or more compoundsof Formula (I).

A sixth aspect of the disclosure relates to a method of treating adisease or disorder associated with reduced nicotinamide adeninedinucleotide (NAD⁺) levels comprising administering to the subjectsuffering from or susceptible to developing a disease or disorderassociated with reduced NAD⁺ levels a therapeutically effective amountof one or more compounds of Formula (I).

A seventh aspect of the disclosure relates to a method of preventing adisease or disorder associated with reduced nicotinamide adeninedinucleotide (NAD⁺) levels comprising administering to the subjectsuffering from or susceptible to developing a disease or disorderassociated with reduced NAD⁺ levels a therapeutically effective amountof one or more compounds of Formula (I).

An eighth aspect of the disclosure relates to a method of reducing therisk of a disease or disorder associated with reduced nicotinamideadenine dinucleotide (NAD⁺) levels comprising administering to thesubject suffering from or susceptible to developing a disease ordisorder associated with reduced NAD⁺ levels a therapeutically effectiveamount of one or more compounds of Formula (I).

An ninth aspect of the disclosure relates to a method of treating adisorder associated with mitochondrial dysfunction comprisingadministering to the subject suffering from or susceptible to developinga metabolic disorder a therapeutically effective amount of one or morecompounds of Formula (I) that increases intracellular nicotinamideadenine dinucleotide (NAD⁺).

A tenth aspect of the disclosure relates to a method of promotingoxidative metabolism comprising administering to the subject sufferingfrom or susceptible to developing a metabolic disorder a therapeuticallyeffective amount of one or more compounds of Formula (I) that increasesintracellular nicotinamide adenine dinucleotide (NAD⁺).

An eleventh aspect of the disclosure relates to a method for themanufacture of a medicament for treating a disease or condition mediatedby ACMSD, wherein the medicament comprises a compound of Formula (I), ora pharmaceutically acceptable salt thereof.

A twelfth aspect of the disclosure relates to a pharmaceuticalcomposition for use in a method for treating a disease or conditionmediated by ACMSD, wherein the medicament comprises a compound ofFormula (I), or a pharmaceutically acceptable salt thereof.

A thirteenth aspect of the disclosure relates to a method of treating adisease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction, comprising administering to a subject in need thereof, atherapeutically effective amount of compound having one of the followingFormulae:

or a pharmaceutically acceptable salt thereof. A fourteenth aspect ofthe disclosure relates to the use of a compound of Formula (I), or apharmaceutically acceptable salt thereof in the manufacture of amedicament for treating, preventing or reducing the risk of a disease ordisorder associated with α-amino-β-carboxymuconate-ε-semialdehydedecarboxylase (ACMSD) dysfunction.

A fifteenth aspect of the disclosure relates to the use of a compound ofFormula (I), or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for treating, preventing or reducing therisk of a disease or disorder associated with reduced nicotinamideadenine dinucleotide (NAD⁺) levels.

A sixteenth aspect of the disclosure relates to the use of a compound ofFormula (I), or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for treating, preventing or reducing therisk of a disorder associated with mitochondrial dysfunction.

A seventeenth aspect of the disclosure relates to the use of a compoundof Formula (I), or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for promoting oxidative metabolism.

A eighteenth aspect of the disclosure relates to the use of a compoundof Formula (I), a pharmaceutically acceptable salt thereof in themanufacture of a medicament for treating, preventing or reducing therisk of a disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction.

A nineteenth aspect of the disclosure relates to a compound of Formula(I), or a pharmaceutically acceptable salt thereof for use as amedicament for treating, preventing or reducing the risk of a disease ordisorder associated with reduced nicotinamide adenine dinucleotide(NAD⁺) levels.

A twentieth aspect of the disclosure relates to a compound of Formula(I), or a pharmaceutically acceptable salt thereof for use as amedicament for treating, preventing or reducing the risk of a disorderassociated with mitochondrial dysfunction.

A twenty first aspect of the disclosure relates to a compound of Formula(I), or a pharmaceutically acceptable salt thereof for use in treating,preventing or reducing the risk of a disease or disorder associated withreduced nicotinamide adenine dinucleotide (NAD⁺) levels.

A twenty second aspect of the disclosure relates to a compound ofFormula (I), or a pharmaceutically acceptable salt thereof for use infor treating, preventing or reducing the risk of a disorder associatedwith mitochondrial dysfunction.

A twenty third aspect of the disclosure relates to a compound of Formula(I), or a pharmaceutically acceptable salt thereof for use in promotingoxidative metabolism.

A twenty fourth aspect of the disclosure relates to a compound havingthe one of the following Formula:

or a pharmaceutically acceptable salt thereof in the manufacture of amedicament for treating a disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction.

A twenty fifth aspect of the disclosure relates to a compound having theone of the following Formula:

or a pharmaceutically acceptable salt thereof for use as a medicamentfor treating a disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction.

A twenty sixth aspect of the disclosure relates to a compound having theone of the following Formula:

or a pharmaceutically acceptable salt thereof

for use in treating a disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction.

In certain aspects, the ACMSD modulating compounds may be administeredalone or in combination with other compounds, including other ACMSDmodulating compounds, or other therapeutic agents.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this disclosure belongs. In the specification, thesingular forms also include the plural unless the context clearlydictates otherwise. Although methods and materials similar to orequivalent to those described herein can be used in the practice andtesting of the disclosure, suitable methods and materials are describedbelow. All publications, patent applications, patents, and otherreferences mentioned herein are incorporated by reference. Thereferences cited herein are not admitted to be prior art to the claimeddisclosure. In the case of conflict, the present specification,including definitions, will control. In addition, the materials,methods, and examples are illustrative only and not intended to belimiting.

Other features and advantages of the disclosure will be apparent fromthe following detailed description and claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph of the measured NAD⁺ levels in human primaryhepatocytes treated with Compound 4, detected by LC-MS/MS.

FIG. 2 is a graph of the measured NAD⁺ levels in murine primaryhepatocytes treated with different concentrations of Compound 17 for 24hours and detected by LC-MS/MS. The data indicate an increase in NAD⁺levels in murine primary hepatocytes treated with Compound 17.

FIG. 3 is a graph of the measured NAD⁺ content in human primaryhepatocytes treated with various concentrations of Compound 1 andmono-(2-ethylhexyl)phthalate (MEHP), as a control.

FIG. 4A is a graph of gene expression of Acmsd, Sod-1, and Sod-2 asdetermined by RT-qPCR in AML-12 cells treated with Compound 1 for 24hours. FIG. 4B is a graph of gene expression of Sod-1 and Sod-2 asdetermined by RT-qPCR in Hepa-1.6 cells treated with Compound 1 for 24hours. FIG. 4C is a graph of gene expression of Acmsd, Sod-1, Sod-2, andPgc1a as determined by RT-qPCR in primary mouse hepatocytes treated withCompound 17 for 24 hours. Bar graphs represent mean±SEM, ***p≦0.005

FIG. 5A is a graph showing the modulation of SOD2 activity in AML-12cells treated for 24 hours with Compound 1. FIG. 5B shows a graph of themodulation of SOD2 activity in AML-12 cells treated for 24 hours withCompound 17. FIG. 5C shows a graph of the modulation of SOD2 activity inprimary murine hepatocytes cells treated for 24 hours with Compound 17.

FIG. 6A is a gel showing the effect of Compound 1 on the FoxO1phosphorylation levels. FIG. 6B is a gel showing the effect of Compound17 on the FoxO1 phosphorylation levels.

FIG. 7A is a graph of changes in acsmd-1 and sod-3 expression at mRNAlevels measured in N2 wild type worms at day 2 of adulthood by acmsd-1RNAi silencing in Caenorhabditis elegans (C. elegans). FIG. 7B is agraph of the induction of sod-3 expression at protein levels in N2 wormsat day 3 of adulthood, quantified by using SOD-3 gfp reporter strain,after acmsd-1 RNAi silencing in C. elegans. FIG. 7C is a graph showingthe survival of worms upon downregulation of acmsd-1 by feeding aspecific RNAi in C. elegans. FIG. 7C shows that downregulation ofacmsd-1 improves the survival of worms in a SIR-2.1 and DAF-16 dependentmanner. FIG. 7D is a graph showing that downregulation of acmsd-1improves the stress-resistance of worms when they are exposed toparaquat-induced oxidative stress. FIG. 7E is a graph showing themobility of worms over time fed with acmsd-1 RNAi under paraquat-inducedoxidative stress condition. As FIGS. 7C-7E show, reduced acmsd-1expression improves the survival and fitness of worms underparaquat-induced oxidative stress. FIG. 7F is a graph that shows thesurvival of worms under paraquat-induced stress conditions when exposedto acmsd-1 RNAi during different stages of development. FIG. 7Fillustrates that the improvement of the survival of worms under paraquatconditions is independent of the developmental stage at which the wormswere exposed to the acmsd-1 RNAi. FIG. 7G is a graph showing wormsurvival under paraquat-induced stress conditions upon downregulation ofacmsd-1 combined with daf-16 downregulation by feeding a specific RNAiin C. elegans. FIG. 7G shows that the improved survival of worms withdownregulated acmsd-1 is daf-16 dependent under paraquat-inducedoxidative stress conditions.

FIG. 8A is a graph of changes in caspase3/7 activity induced bycisplatin in MDCK cells when treated with different concentrations ofCompound. 18 in combination with cisplatin. FIG. 8B is a graph ofchanges in caspase3/7 activity induced by cisplatin in MDCK cells whentreated with different concentrations of Compound. 18 one hour prior tothe addition of cisplatin.

DETAILED DESCRIPTION OF THE DISCLOSURE

Compounds of Formula (I)

The present disclosure relates to compounds of Formula (I):

or a pharmaceutically acceptable salt or tautomer thereof,

wherein:

X is O, OH, or Cl;

L is —(CH₂)_(m)CH₂CH₂—, —(CH₂)_(m)Y(CH₂)_(p)—, —(CH₂)_(m)C(O)(CH₂)_(p)—,—(CH₂)_(m)C(O)O(CH₂)_(p)—, —(CH₂)_(m)C(O)NR²(CH₂)_(p)—, or—(CH₂)_(m)NR²C(O)(CH₂)_(p);

Y is O, N or S(O)_(q);

R¹ is C₆-C₁₀ aryl or heteroaryl, wherein the aryl and heteroaryl aresubstituted with R^(a) and R^(b), and optionally substituted with one ormore R^(e);

R² is H or C₁-C₆ alkyl;

one of R^(a) and R^(b) is hydrogen and the other is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)tetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r) isoxazol-3-ol,—(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH, —(CH₂)_(r)C(O)NHCN, or—(CH₂)_(r)C(O)NHS(O)₂alkyl;

R^(c) is H, C₁-C₆ alkyl, C₁-C₆ haloalkyl, halogen, —CN, —OR^(x),—CO₂R^(x), or NO₂;

R^(d) is methyl, optionally substituted 5- to 10-membered aryl,optionally substituted 5- or 6-membered heteroaryl, or optionallysubstituted 5- or 6-membered carbocycle;

each R^(x) is independently at each occurrence hydrogen or C₁-C₆ alkyl;

each R^(e) is independently C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,halogen, —OR^(y), C₁-C₆ haloalkyl, —NHR^(z), —OH, or —CN;

R^(f) is H or absent;

each R^(y) and R^(z) is independently hydrogen, C₁-C₆ alkyl, or C₁-C₆haloalkyl;

each m and p independently is 0, 1 or 2, wherein m+p<3;

q is 0, 1, or 2;

r is 0 or 1; and

the dotted line is an optional double bond;

with the proviso that R^(c) is not hydrogen or —CN when X is O, L is—SCH₂— and R^(d) is optionally substituted phenyl, R^(c) is not C₁-C₆alkyl when X is O, L is —SCH₂— and R^(d) is methyl, and that R^(c) isnot —CN when L is —SCH₂— and R^(d) is 2-furyl.

In some embodiments of Formula (I), X is O, OH, or Cl. In otherembodiments, X is O. In other embodiments, X is OH. In otherembodiments, X is Cl.

In some embodiments of Formula (I), L is —(CH₂)_(m)CH₂CH₂—,—(CH₂)_(m)Y(CH₂)_(p)—, —(CH₂)_(m)C(O)(CH₂)_(p)—,—(CH₂)_(m)C(O)O(CH₂)_(p)—, —(CH₂)_(m)C(O)NR²(CH₂)_(p)—, or—(CH₂)_(m)NR²C(O)(CH₂)_(p). In other embodiments, L is —CH₂CH₂—,—CH₂CH₂CH₂—, —SCH₂—, —SCH₂CH₂—, —CH₂S—, —CH₂SCH₂—, —CH₂CH₂S—, —S(O)CH₂—,—S(O)CH₂CH₂—, —CH₂S(O)—, —CH₂S(O)CH₂—, —CH₂CH₂S(O)—, —S(O)₂CH₂—,—S(O)₂CH₂CH₂—, —CH₂S(O)₂—, —CH₂S(O)₂CH₂—, —CH₂CH₂S(O)₂—, —OCH₂—,—OCH₂CH₂—, —CH₂O—, —CH₂OCH₂—, —CH₂CH₂O—, —NR²CH₂—, —CH₂NR²—,—CH₂NR²CH₂—, —CH₂CH₂NR²—, —NR²CH₂CH₂—, —C(O)CH₂—, —C(O)CH₂CH₂—, —C(O)O—,—C(O)OCH₂—, —CH₂C(O)O—, —C(O)NR²—, —C(O)NR²CH₂—, —NR²C(O), —NR²C(O)CH₂,or —CH₂NR²C(O). In other embodiments, L is —CH₂CH₂—, —CH₂CH₂CH₂—,—SCH₂—, —SCH₂CH₂—, —S(O)CH₂—, —S(O)CH₂CH₂—, —S(O)₂CH₂—, —S(O)₂CH₂CH₂—,—OCH₂—, —OCH₂CH₂—, —NR²CH₂—, —NR²CH₂CH₂—, —C(O)CH₂—, —C(O)CH₂CH₂—,—C(O)O—, —C(O)OCH₂—, —CH₂C(O)O—, —C(O)NR²—, —C(O)NR²CH₂—, —NR²C(O), or—NR²C(O)CH₂. In other embodiments, L is —CH₂CH₂—, —CH₂C(O)—, —C(O)CH₂—,—NR²CH₂—, —CH₂N^(R2)—, —OCH₂—, —CH₂O—, —SCH₂—, —CH₂S—, —S(O)CH₂—,—CH₂S(O)—, —CH₂S(O)₂—, or —S(O)₂CH₂—.

In some embodiments of Formula (I), R¹ is C₆-C₁₀ aryl or heteroaryl,wherein the aryl and heteroaryl are substituted with R^(a) and R^(b),and optionally substituted with one or more R^(e). In other embodiments,R¹ is C₆-C₁₀ aryl substituted with R^(a) and R^(b), and optionallysubstituted with one or more R^(e). In other embodiments, R¹ isheteroaryl substituted with R^(a) and R^(b), and optionally substitutedwith one or more R^(e). In further embodiments, R¹ is phenyl substitutedwith R^(a) and R^(b), and optionally substituted with one or more R^(e).

In some embodiments of Formula (I), R^(a) is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)tetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r) isoxazol-3-ol,—(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH, —(CH₂)_(r)C(O)NHCN, or—(CH₂)_(r)C(O)NHS(O)₂alkyl. In other embodiments, R^(a) is—(CH₂)_(r)CO₂R^(x), —OCH₂CO₂R^(x), tetrazole, —(CH₂)tetrazole,oxadiazolone, —(CH₂)oxadiazolone, tetrazolone, —(CH₂)tetrazolone,thiadiazolol, —(CH₂)thiadiazolol, isoxazol-3-ol, —(CH₂) isoxazol-3-ol,—P(O)(OH)OR^(x), —(CH₂)P(O)(OH)OR^(x), —S(O)₂OH, —(CH₂)S(O)₂OH,—C(O)NHCN —(CH₂)C(O)NHCN, —C(O)NHS(O)₂alkyl, or —(CH₂)C(O)NHS(O)₂alkyl.In other embodiments, R^(a) is hydrogen, CO₂R^(x), CH₂CO₂R^(x),tetrazole, or oxadiazolone. In further embodiments, R^(a) is hydrogen,CO₂H, CH₂CO₂H, tetrazole, or 1,2,4-oxadiazol-5(4H)-one.

In some embodiments of Formula (I), R^(b) is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)tetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r) isoxazol-3-ol,—(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH, —(CH₂)_(r)C(O)NHCN, or—(CH₂)_(r)C(O)NHS(O)₂alkyl. In other embodiments, R^(b) is—(CH₂)_(r)CO₂R^(x), —OCH₂CO₂R^(x), tetrazole, —(CH₂)tetrazole,oxadiazolone, —(CH₂)oxadiazolone, tetrazolone, —(CH₂)tetrazolone,thiadiazolol, —(CH₂)thiadiazolol, isoxazol-3-ol, —(CH₂) isoxazol-3-ol,—P(O)(OH)OR^(x), —(CH₂)P(O)(OH)OR^(x), —S(O)₂OH, —(CH₂)S(O)₂OH,—C(O)NHCN —(CH₂)C(O)NHCN, —C(O)NHS(O)₂alkyl, or —(CH₂)C(O)NHS(O)₂alkyl.In other embodiments, R^(b) is hydrogen, CO₂R^(x), CH₂CO₂R^(x),tetrazole, or oxadiazolone. In further embodiments, R^(b) is hydrogen,CO₂H, CH₂CO₂H, tetrazole, or 1,2,4-oxadiazol-5(4H)-one. In furtherembodiments, R^(b) is hydrogen.

In some embodiments of Formula (I), R^(c) is H, C₁-C₆ alkyl, C₁-C₆haloalkyl, halogen, —CN, —OR^(x), —CO₂R^(x), or NO₂. In otherembodiments, R^(c) is C₁-C₆ alkyl, C₁-C₆ haloalkyl, halogen, —CN,—OR^(x), —CO₂R^(x), or NO₂. In other embodiments, R^(c) is halogen, —CN,—OR^(x), or C₁-C₆ alkyl. In other embodiments, R^(c) is halogen, —CN,—OR^(x), or C₁-C₃ alkyl. In other embodiments, R^(c) is H, —CN, orhalogen. In other embodiments, R^(c) is —CN or halogen.

In some embodiments of Formula (I), R^(d) is methyl, optionallysubstituted 5- to 10-membered aryl, optionally substituted 5- or6-membered heteroaryl, or optionally substituted 5- or 6-memberedcarbocycle. In other embodiments, R^(d) is methyl, optionallycyclohexyl, optionally substituted pyridinyl, optionally substitutedthiazolyl, optionally substituted phenyl, or optionally substitutedthienyl. In other embodiments, R^(d) is methyl, cyclohexyl, pyridinyl,thiazolyl, phenyl, or thienyl. In other embodiments, R^(d) iscyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each isoptionally substituted with one or more substituents independentlyselected from halogen, C₁-C₆ alkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy,C₁-C₆ haloalkyl, C₁-C₆ haloalkoxy, —OH, CN, and amino. In otherembodiments, R^(d) is cyclohexyl, pyridinyl, thiazolyl, phenyl, orthienyl, wherein each is optionally substituted with one or moresubstituents independently selected from halogen, C₁-C₆ alkyl, C₁-C₆hydroxyalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, and C₁-C₆ haloalkoxy. Inother embodiments, R^(d) is cyclohexyl, pyridinyl, thiazolyl, phenyl, orthienyl, wherein each is optionally substituted with one or morehalogen. In yet other embodiments, R^(d) is cyclohexyl, pyridinyl,thiazolyl, phenyl, or thienyl. In other embodiments, R^(d) iscyclohexyl, pyridinyl, thiazolyl, phenyl, 4-chlorophenyl,4-methylphenyl, or thienyl.

In some embodiments of Formula (I), each R^(e) is independently C₁-C₆alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, halogen, —OR^(y), C₁-C₆ haloalkyl,—NHR^(z), —OH, or —CN. In other embodiments, C₁-C₄ alkyl, C₂-C₄ alkenyl,C₂-C₄ alkynyl, halogen, —OR^(y), C₁-C₄ haloalkyl, —NHR^(z), —OH, or —CN.

In some embodiments of Formula (I), R^(f) is H or absent. In otherembodiments, R^(f) is H. In other embodiments, R^(f) is absent, when Nto which it is attached participates in a double bond.

In some embodiments of Formula (I), R^(x) is hydrogen or C₁-C₆ alkyl. Inother embodiments, R^(x) is hydrogen or C₁-C₃ alkyl. In furtherembodiments, R^(x) is hydrogen, methyl, ethyl, n-propyl, or iso-propyl.

In some embodiments of Formula (I), R^(y) is independently hydrogen,C₁-C₆ alkyl, or C₁-C₆ haloalkyl. In other embodiments, R^(y) ishydrogen, C₁-C₃ alkyl, or C₁-C₃ haloalkyl.

In some embodiments of Formula (I), each R^(z) is independentlyhydrogen, C₁-C₆ alkyl, or C₁-C₆ haloalkyl. In other embodiments, R^(z)is hydrogen, C₁-C₃ alkyl, or C₁-C₃ haloalkyl.

In some embodiments of Formula (I), m is 0, 1 or 2. In otherembodiments, m is 0. In other embodiments, m is 1. In yet otherembodiments, m is 2.

In some embodiments of Formula (I), p is 0, 1 or 2. In otherembodiments, p is 0. In other embodiments, p is 1. In yet otherembodiments, p is 2.

In some embodiments of Formula (I), m+p<3;

In some embodiments of Formula (I), q is 0, 1, or 2. In otherembodiments, q is 0. In other embodiments, q is 1. In other embodiments,q is 2.

In some embodiments of Formula (I), r is 0 or 1. In other embodiments, ris 0. In other embodiments, r is 1.

In some embodiments of Formula (I), the dotted line is a single bond. Inother embodiments, the dotted line is a double bond.

In some embodiments of Formula (I), one of R^(a) and R^(b) is hydrogenand the other is CO₂R^(x), CH₂CO₂R^(x), tetrazole, or oxadiazolone. Inother embodiments, R^(b) is hydrogen and R^(a) is CH₂CO₂H, tetrazole, or(1,2,4-oxadiazol-5(4H)-one).

In some embodiments of Formula (I), R^(b) is hydrogen, R^(c) is —CN,R^(d) is thienyl, and R^(a) is CH₂CO₂H, tetrazole, or(1,2,4-oxadiazol-5(4H)-one).

In some embodiments of Formula (I), R^(c) is halogen, R^(a) is —CO₂H,and R^(b) is H. In other embodiments, R^(c) is —Br, R^(a) is —CO₂H, andR^(b) is H. In further embodiments, R^(c) is —Cl, R^(a) is —CO₂H, andR^(b) is H.

In some embodiments of Formula (I), R^(c) is halogen, R^(a) istetrazole, and R^(b) is H. In other embodiments, R^(c) is —Br, R^(a) istetrazole, and R^(b) is H. In further embodiments, R^(c) is —Cl, R^(a)is tetrazole, and R^(b) is H.

In some embodiments of Formula (I), R^(c) is halogen, R^(a) is —CH₂CO₂H,and R^(b) is H. In other embodiments, R^(c) is —Br, R^(a) is —CH₂CO₂H,and R^(b) is H. In further embodiments, R^(c) is —Cl, R^(a) is —CH₂CO₂H,and R^(b) is H.

In some embodiments of Formula (I), R^(c) is halogen, R^(a) is(1,2,4-oxadiazol-5(4H)-one), and R^(b) is H. In other embodiments, R^(c)is —Br, R^(a) is (1,2,4-oxadiazol-5(4H)-one), and R^(b) is H. In otherembodiments, R^(c) is —Cl, R^(a) is (1,2,4-oxadiazol-5(4H)-one), andR^(b) is H.

In some embodiments of Formula (I), R^(c) is —CN, R^(a) is —CO₂H, andR^(b) is H. In other embodiments, R^(c) is —CN, R^(a) is —CH₂CO₂H, andR^(b) is H. In other embodiments, R^(c) is —CN, R^(a) is tetrazole, andR^(b) is H. In yet other embodiments, R^(c) is —CN, R^(a) is(1,2,4-oxadiazol-5(4H)-one), and R^(b) is H.

In some embodiments of Formula (I), R^(c) is not hydrogen or —CN and Xis O, L is —SCH₂— and R^(d) is optionally substituted phenyl. In otherembodiments, R^(c) is not C₁-C₆ alkyl and X is O, L is —SCH₂— and R^(d)is methyl. In other embodiments, R^(c) is not —CN and X is O, L is—SCH₂— and R^(d) is 2-furyl.

In some embodiments of Formula (I), R^(c) is not hydrogen or —CN when Xis O, L is —SCH₂— and R^(d) is optionally substituted phenyl.

In some embodiments of Formula (I), R^(c) is not C₁-C₆ alkyl when X isO, L is —SCH₂— and R^(d) is methyl.

In some embodiments of Formula (I), R^(c) is not —CN when X is O, L is—SCH₂— and R^(d) is 2-furyl.

In one embodiment, the compound of Formula (I) is represented by Formula(Ia):

or a pharmaceutically acceptable salt, or tautomer thereof,

wherein:

L is —(CH₂)_(m)CH₂CH₂—, —(CH₂)_(m)Y(CH₂)_(p)—, —(CH₂)_(m)C(O)(CH₂)_(p)—,—(CH₂)_(m)C(O)O(CH₂)_(p)—, —(CH₂)_(m)C(O)NR²(CH₂)_(p)—, or—(CH₂)_(m)NR²C(O)(CH₂)_(p);

Y is O, N or S(O)_(q);

R¹ is C₆-C₁₀ aryl or heteroaryl, wherein the aryl and heteroaryl aresubstituted with R^(a) and R^(b), and optionally substituted with one ormore R^(e);

R² is H or C₁-C₆ alkyl;

one of R^(a) and R^(b) is hydrogen and the other is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)tetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r) isoxazol-3-ol,—(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH, —(CH₂)_(r)C(O)NHCN, or—(CH₂)_(r)C(O)NHS(O)₂alkyl;

R^(c) is H, C₁-C₆ alkyl, C₁-C₆ haloalkyl, halogen, —CN, —OR^(y),—CO₂R^(x), or NO₂;

R^(d) is methyl, optionally substituted 5- to 10-membered aryl,optionally substituted 5- or 6-membered heteroaryl, or optionallysubstituted 5- or 6-membered carbocycle;

each R^(x) is independently at each occurrence hydrogen or C₁-C₆ alkyl;

each R^(e) is independently C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,halogen, —OR^(y), C₁-C₆ haloalkyl, —NHR^(z), —OH, or —CN;

each R^(y) and R^(z) is independently hydrogen, C₁-C₆ alkyl, or C₁-C₆haloalkyl;

each m and p independently is 0, 1 or 2, wherein m+p<3;

q is 0, 1, or 2; and

r is 0 or 1;

with the proviso that R^(c) is not hydrogen or —CN when L is —SCH₂— andR^(d) is optionally substituted phenyl, R^(c) is not C₁-C₆ alkyl when Lis —SCH₂— and R^(d) is methyl, and that R^(c) is not —CN when L is—SCH₂— and R^(d) is 2-furyl.

In some embodiments of Formula (Ia),

L is —CH₂CH₂—, —CH₂C(O)—, —C(O)CH₂—, —NR²CH₂—, —CH₂NR²—, —OCH₂—, —CH₂O—,—SCH₂—, —CH₂S—, —S(O)CH₂—, —CH₂S(O)—, —CH₂S(O)₂—, or —S(O)₂CH₂;

Y is O, N or S(O)_(q);

R¹ is C₆-C₁₀ aryl or heteroaryl, wherein the aryl and heteroaryl aresubstituted with R^(a) and R^(b), and optionally substituted with one ormore R^(e);

R² is H or C₁-C₆ alkyl;

one of R^(a) and R^(b) is hydrogen and the other is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)tetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r) isoxazol-3-ol,—(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH, —(CH₂)_(r)C(O)NHCN, or—(CH₂)_(r) C(O)NHS(O)₂alkyl;

R^(c) is C₁-C₆ alkyl, C₁-C₆ haloalkyl, halogen, —CN, —OR^(x), —CO₂R^(x),or NO₂;

R^(d) is methyl, optionally substituted 5- to 10-membered aryl,optionally substituted 5- or 6-membered heteroaryl, or optionallysubstituted 5- or 6-membered carbocycle;

each R^(x) is independently at each occurrence hydrogen or C₁-C₆ alkyl;

each R^(e) is independently C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,halogen, —OR^(y), C₁-C₆ haloalkyl, —NHR^(z), —OH, or —CN;

each R^(y) and R^(z) is independently hydrogen, C₁-C₆ alkyl, or C₁-C₆haloalkyl;

each m and p independently is 0, 1 or 2, wherein m+p<3;

q is 0, 1, or 2; and

r is 0 or 1;

with the proviso that R^(c) is not —CN when L is —SCH₂— and R^(d) isoptionally substituted phenyl, R^(c) is not C₁-C₆ alkyl when L is —SCH₂—and R^(d) is methyl, and that R^(c) is not —CN when L is —SCH₂— andR^(d) is 2-furyl.

In some embodiments of Formula (Ia), L is —(CH₂)_(m)CH₂CH₂—,—(CH₂)_(m)Y(CH₂)_(p)—, —(CH₂)_(m)C(O)(CH₂)_(p)—,—(CH₂)_(m)C(O)O(CH₂)_(p)—, —(CH₂)_(m)C(O)NR²(CH₂)_(p)—, or—(CH₂)_(m)NR²C(O)(CH₂)_(p). In other embodiments, L is —CH₂CH₂—,—CH₂CH₂CH₂—, —SCH₂—, —SCH₂CH₂—, —CH₂S—, —CH₂SCH₂—, —CH₂CH₂S—, —S(O)CH₂—,—S(O)CH₂CH₂—, —CH₂S(O)—, —CH₂S(O)CH₂—, —CH₂CH₂S(O)—, —S(O)₂CH₂—,—S(O)₂CH₂CH₂—, —CH₂S(O)₂—, —CH₂S(O)₂CH₂—, —CH₂CH₂S(O)₂—, —OCH₂—,—OCH₂CH₂—, —CH₂O—, —CH₂OCH₂—, —CH₂CH₂O—, —NR²CH₂—, —CH₂NR²—,—CH₂NR²CH₂—, —CH₂CH₂NR²—, —NR²CH₂CH₂—, —C(O)CH₂—, —C(O)CH₂CH₂—, —C(O)O—,—C(O)OCH₂—, —CH₂C(O)O—, —C(O)NR²—, —C(O)NR²CH₂—, —NR²C(O), —NR²C(O)CH₂,or —CH₂NR₂C(O). In other embodiments, L is —CH₂CH₂—, —CH₂CH₂CH₂—,—SCH₂—, —SCH₂CH₂—, —S(O)CH₂—, —S(O)CH₂CH₂—, —S(O)₂CH₂—, —S(O)₂CH₂CH₂—,—OCH₂—, —OCH₂CH₂—, —NR²CH₂—, —NR²CH₂CH₂—, —C(O)CH₂—, —C(O)CH₂CH₂—,—C(O)O—, —C(O)OCH₂—, —CH₂C(O)O—, —C(O)NR²—, —C(O)NR²CH₂—, —NR²C(O), or—NR²C(O)CH₂. In other embodiments, L is —CH₂CH₂—, —CH₂C(O)—, —C(O)CH₂—,—NR²CH₂—, —CH₂NR²—, —OCH₂—, —CH₂O—, —SCH₂—, —CH₂S—, —S(O)CH₂—,—CH₂S(O)—, —CH₂S(O)₂—, or —S(O)₂CH₂—.

In some embodiments of Formula (Ia), R¹ is C₆-C₁₀ aryl or heteroaryl,wherein the aryl and heteroaryl are substituted with R^(a) and R^(b),and optionally substituted with one or more R^(e). In other embodiments,R¹ is C₆-C₁₀ aryl substituted with R^(a) and R^(b), and optionallysubstituted with one or more R^(e). In other embodiments, R¹ isheteroaryl substituted with R^(a) and R^(b), and optionally substitutedwith one or more R^(e). In further embodiments, R¹ is phenyl substitutedwith R^(a) and R^(b), and optionally substituted with one or more R^(e).

In some embodiments of Formula (Ia), R^(a) is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)tetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r) isoxazol-3-ol,—(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH, —(CH₂)_(r)C(O)NHCN, or—(CH₂)_(r)C(O)NHS(O)₂alkyl. In other embodiments, R^(a) is—(CH₂)_(r)CO₂R^(x), —OCH₂CO₂R^(x), tetrazole, —(CH₂)tetrazole,oxadiazolone, —(CH₂)oxadiazolone, tetrazolone, —(CH₂)tetrazolone,thiadiazolol, —(CH₂)thiadiazolol, isoxazol-3-ol, —(CH₂) isoxazol-3-ol,—P(O)(OH)OR^(x), —(CH₂)P(O)(OH)OR^(x), —S(O)₂OH, —(CH₂)S(O)₂OH,—C(O)NHCN —(CH₂)C(O)NHCN, —C(O)NHS(O)₂alkyl, or —(CH₂)C(O)NHS(O)₂alkyl.In other embodiments, R^(a) is hydrogen, CO₂R^(x), CH₂CO₂R^(x),tetrazole, or oxadiazolone. In further embodiments, R^(a) is hydrogen,CO₂H, CH₂CO₂H, tetrazole, or 1,2,4-oxadiazol-5(4H)-one.

In some embodiments of Formula (Ia), R^(b) is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)tetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r) isoxazol-3-ol,—(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH, —(CH₂)_(r)C(O)NHCN, or—(CH₂)_(r)C(O)NHS(O)₂alkyl. In other embodiments, R^(b) is—(CH₂)_(r)CO₂R^(x), —OCH₂CO₂R^(x), tetrazole, —(CH₂)tetrazole,oxadiazolone, —(CH₂)oxadiazolone, tetrazolone, —(CH₂)tetrazolone,thiadiazolol, —(CH₂)thiadiazolol, isoxazol-3-ol, —(CH₂) isoxazol-3-ol,—P(O)(OH)OR^(x), —(CH₂)P(O)(OH)OR^(x), —S(O)₂OH, —(CH₂)S(O)₂OH,—C(O)NHCN —(CH₂)C(O)NHCN, —C(O)NHS(O)₂alkyl, or —(CH₂)C(O)NHS(O)₂alkyl.In other embodiments, R^(b) is hydrogen, CO₂R^(x), CH₂CO₂R^(x),tetrazole, or oxadiazolone. In further embodiments, R^(b) is hydrogen,CO₂H, CH₂CO₂H, tetrazole, or 1,2,4-oxadiazol-5(4H)-one. In furtherembodiments, R^(b) is hydrogen.

In some embodiments of Formula (Ia), R^(c) is H, C₁-C₆ alkyl, C₁-C₆haloalkyl, halogen, —CN, —OR^(x), —CO₂R^(x), or NO₂. In otherembodiments, R^(c) is C₁-C₆ alkyl, C₁-C₆ haloalkyl, halogen, —CN,—OR^(x), —CO₂R^(x), or NO₂. In other embodiments, R^(c) is halogen, —CN,—OR^(x), or C₁-C₆ alkyl. In other embodiments, R^(c) is halogen, —CN,—OR^(x), or C₁-C₃ alkyl. In other embodiments, R^(c) is H, —CN, orhalogen. In other embodiments, R^(c) is —CN or halogen.

In some embodiments of Formula (Ia), R^(d) is methyl, optionallysubstituted 5- to 10-membered aryl, optionally substituted 5- or6-membered heteroaryl, or optionally substituted 5- or 6-memberedcarbocycle. In other embodiments, R^(d) is methyl, optionallycyclohexyl, optionally substituted pyridinyl, optionally substitutedthiazolyl, optionally substituted phenyl, or optionally substitutedthienyl. In other embodiments, R^(d) is cyclohexyl, pyridinyl,thiazolyl, phenyl, or thienyl, wherein each is optionally substitutedwith one or more substituents independently selected from halogen, C₁-C₆alkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, C₁-C₆haloalkoxy, —OH, CN, and amino. In other embodiments, R^(d) iscyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each isoptionally substituted with one or more substituents independentlyselected from halogen, C₁-C₆ alkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy,C₁-C₆ haloalkyl, and C₁-C₆ haloalkoxy. In other embodiments, R^(d) iscyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each isoptionally substituted with one or more halogen. In other embodiments,R^(d) is methyl, cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl.In yet other embodiments, R^(d) is cyclohexyl, pyridinyl, thiazolyl,phenyl, or thienyl. In other embodiments, R^(d) is cyclohexyl,pyridinyl, thiazolyl, phenyl, 4-chlorophenyl, 4-methylphenyl, orthienyl.

In some embodiments of Formula (Ia), each R^(e) is independently C₁-C₆alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, halogen, —OR^(y), C₁-C₆ haloalkyl,—NHR^(z), —OH, or —CN. In other embodiments, C₁-C₄ alkyl, C₂-C₄ alkenyl,C₂-C₄ alkynyl, halogen, —OR^(y), C₁-C₄ haloalkyl, —NHR^(z), —OH, or —CN.

In some embodiments of Formula (Ia), R^(x) is hydrogen or C₁-C₆ alkyl.In other embodiments, R^(x) is hydrogen or C₁-C₃ alkyl. In furtherembodiments, R^(x) is hydrogen, methyl, ethyl, n-propyl, or iso-propyl.

In some embodiments of Formula (Ia), R^(y) is independently hydrogen,C₁-C₆ alkyl, or C₁-C₆ haloalkyl. In other embodiments, R^(y) ishydrogen, C₁-C₃ alkyl, or C₁-C₃ haloalkyl.

In some embodiments of Formula (Ia), each R^(z) is independentlyhydrogen, C₁-C₆ alkyl, or C₁-C₆ haloalkyl. In other embodiments, R^(z)is hydrogen, C₁-C₃ alkyl, or C₁-C₃ haloalkyl.

In some embodiments of Formula (Ia), m is 0, 1 or 2. In otherembodiments, m is 0. In other embodiments, m is 1. In yet otherembodiments, m is 2.

In some embodiments of Formula (Ia), p is 0, 1 or 2. In otherembodiments, p is 0. In other embodiments, p is 1. In yet otherembodiments, p is 2.

In some embodiments of Formula (Ia), q is 0, 1, or 2. In otherembodiments, q is 0. In other embodiments, q is 1. In other embodiments,q is 2.

In some embodiments of Formula (Ia), r is 0 or 1. In other embodiments,r is 0. In other embodiments, r is 1.

In some embodiments of Formula (Ia), one of R^(a) and R^(b) is hydrogenand the other is CO₂R^(x), CH₂CO₂R^(x), tetrazole, or oxadiazolone. Inother embodiments, R^(b) is hydrogen and R^(a) is CH₂CO₂H, tetrazole, or(1,2,4-oxadiazol-5(4H)-one).

In some embodiments of Formula (Ia), R^(b) is hydrogen, R^(c) is —CN,R^(d) is thienyl, and R^(a) is CH₂CO₂H, tetrazole, or(1,2,4-oxadiazol-5(4H)-one).

In some embodiments of Formula (Ia), R^(c) is halogen, R^(a) is —CO₂H,and R^(b) is H. In other embodiments, R^(c) is —Br, R^(a) is —CO₂H, andR^(b) is H. In further embodiments, R^(c) is —Cl, R^(a) is —CO₂H, andR^(b) is H.

In some embodiments of Formula (Ia), R^(c) is halogen, R^(a) istetrazole, and R^(b) is H. In other embodiments, R^(c) is —Br, R^(a) istetrazole, and R^(b) is H. In further embodiments, R^(c) is —Cl, R^(a)is tetrazole, and R^(b) is H.

In some embodiments of Formula (Ia), R^(c) is halogen, R^(a) is—CH₂CO₂H, and R^(b) is H. In other embodiments, R^(c) is —Br, R^(a) is—CH₂CO₂H, and R^(b) is H. In further embodiments, R^(c) is —Cl, R^(a) is—CH₂CO₂H, and R^(b) is H.

In some embodiments of Formula (Ia), R^(c) is halogen, R^(a) is(1,2,4-oxadiazol-5(4H)-one), and R^(b) is H. In other embodiments, R^(c)is —Br, R^(a) is (1,2,4-oxadiazol-5(4H)-one), and R^(b) is H. In otherembodiments, R^(c) is —Cl, R^(a) is (1,2,4-oxadiazol-5(4H)-one), andR^(b) is H.

In some embodiments of Formula (Ia), R^(c) is —CN, R^(a) is —CO₂H, andR^(b) is H. In other embodiments, R^(c) is —CN, R^(a) is —CH₂CO₂H, andR^(b) is H. In other embodiments, R^(c) is —CN, R^(a) is tetrazole, andR^(b) is H. In yet other embodiments, R^(c) is —CN, R^(a) is(1,2,4-oxadiazol-5(4H)-one), and R^(b) is H.

In some embodiments of Formula (Ia), R^(c) is not hydrogen or —CN and Lis —SCH₂— and R^(d) is optionally substituted phenyl. In otherembodiments, R^(c) is not C₁-C₆ alkyl and L is —SCH₂— and R^(d) ismethyl. In other embodiments, R^(c) is not —CN and L is —SCH₂— and R^(d)is 2-furyl.

In some embodiments of Formula (Ia), R^(c) is not hydrogen or —CN when Lis —SCH₂— and R^(d) is optionally substituted phenyl.

In some embodiments of Formula (Ia), R^(c) is not C₁-C₆ alkyl when L is—SCH₂— and R^(d) is methyl.

In some embodiments of Formula (Ia), R^(c) is not —CN when L is —SCH₂—and R^(d) is 2-furyl.

In another embodiment, the compound of Formula (I) is represented byFormula (Ib):

or a pharmaceutically acceptable salt thereof

wherein:

R^(a) and R^(b) is hydrogen and the other is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)tetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r) isoxazol-3-ol,—(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH, —(CH₂)_(r)C(O)NHCN, or—(CH₂)_(r)C(O)NHS(O)₂alkyl;

R^(c) is H, C₁-C₆ alkyl, C₁-C₆ haloalkyl, halogen, —CN, —OR^(x),—CO₂R^(x), or NO₂;

R^(d) is methyl, optionally substituted 5- to 10-membered aryl,optionally substituted 5- or 6-membered heteroaryl, or optionallysubstituted 5- or 6-membered carbocycle;

each R^(x) is independently at each occurrence hydrogen or C₁-C₆ alkyl;

each R^(e) is independently C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,halogen, —OR^(y), C₁-C₆ haloalkyl, —NHR^(z), —OH, or —CN;

each R^(y) and R^(z) is independently hydrogen, C₁-C₆ alkyl, or C₁-C₆haloalkyl; and

n is 0, 1, 2, or 3;

with the proviso that R^(c) is not hydrogen or —CN when and R^(d) isoptionally substituted phenyl, R^(c) is not C₁-C₆ alkyl when R^(d) ismethyl, and that R^(c) is not —CN when R^(d) is 2-furyl.

In some embodiments of Formula (Ib),

one of R^(a) and R^(b) is hydrogen and the other is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)tetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r) isoxazol-3-ol,—(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH, —(CH₂)_(r)C(O)NHCN, or—(CH₂)_(r) C(O)NHS(O)₂alkyl;

R^(c) is C₁-C₆ alkyl, C₁-C₆ haloalkyl, halogen, —CN, —OR^(x), —CO₂R^(x),or NO₂;

R^(d) is methyl, optionally substituted 5- to 10-membered aryl,optionally substituted 5- or 6-membered heteroaryl, or optionallysubstituted 5- or 6-membered carbocycle;

each R^(x) is independently at each occurrence hydrogen or C₁-C₆ alkyl;

each R^(e) is independently C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,halogen, —OR^(y), C₁-C₆ haloalkyl, —NHR^(z), —OH, or —CN;

with the proviso that R^(c) is not hydrogen or —CN when R^(d) isoptionally substituted phenyl, R^(c) is not C₁-C₆ alkyl when R^(d) ismethyl, and that R^(c) is not —CN when R^(d) is 2-furyl.

In some embodiments of formula (Ib),

one of R^(a) and R^(b) is hydrogen and the other is CO₂R^(x),CH₂CO₂R^(x), tetrazole, or oxadiazolone;

R^(c) is halogen, —CN, —OR^(x), or C₁-C₆ alkyl;

R^(d) is methyl, optionally substituted 5- to 10-membered aryl,optionally substituted 5- or 6-membered heteroaryl, or optionallysubstituted 5- or 6-membered carbocycle; and

R^(x) is hydrogen or C₁-C₆ alkyl;

each R^(e) is independently C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,halogen, —OR^(y), C₁-C₆ haloalkyl, —NHR^(z), —OH, or —CN;

each R^(y) and R^(z) is independently hydrogen, C₁-C₆ alkyl, or C₁-C₆haloalkyl; and

n is 0, 1, 2, or 3;

with the proviso that R^(c) is not —CN when R^(d) is optionallysubstituted phenyl, R^(c) is not C₁-C₆ alkyl when R^(d) is methyl, andthat R^(c) is not —CN when R^(d) is 2-furyl.

In some embodiments of Formula (Ib), R^(a) is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)tetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r) isoxazol-3-ol,—(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH, —(CH₂)_(r)C(O)NHCN, or—(CH₂)_(r)C(O)NHS(O)₂alkyl. In other embodiments, R^(a) is—(CH₂)_(r)CO₂R^(x), —OCH₂CO₂R^(x), tetrazole, —(CH₂)tetrazole,oxadiazolone, —(CH₂)oxadiazolone, tetrazolone, —(CH₂)tetrazolone,thiadiazolol, —(CH₂)thiadiazolol, isoxazol-3-ol, —(CH₂) isoxazol-3-ol,—P(O)(OH)OR^(x), —(CH₂)P(O)(OH)OR^(x), —S(O)₂OH, —(CH₂)S(O)₂OH,—C(O)NHCN —(CH₂)C(O)NHCN, —C(O)NHS(O)₂alkyl, or —(CH₂)C(O)NHS(O)₂alkyl.In other embodiments, R^(a) is hydrogen, CO₂R^(x), CH₂CO₂R^(x),tetrazole, or oxadiazolone. In further embodiments, R^(a) is hydrogen,CO₂H, CH₂CO₂H, tetrazole, or 1,2,4-oxadiazol-5(4H)-one.

In some embodiments of Formula (Ib), R^(b) is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)tetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r) isoxazol-3-ol,—(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH, —(CH₂)_(r)C(O)NHCN, or—(CH₂)_(r)C(O)NHS(O)₂alkyl. In other embodiments, R^(b) is—(CH₂)_(r)CO₂R^(x), —OCH₂CO₂R^(x), tetrazole, —(CH₂)tetrazole,oxadiazolone, —(CH₂)oxadiazolone, tetrazolone, —(CH₂)tetrazolone,thiadiazolol, —(CH₂)thiadiazolol, isoxazol-3-ol, —(CH₂) isoxazol-3-ol,—P(O)(OH)OR^(x), —(CH₂)P(O)(OH)OR^(x), —S(O)₂OH, —(CH₂)S(O)₂OH,—C(O)NHCN —(CH₂)C(O)NHCN, —C(O)NHS(O)₂alkyl, or —(CH₂)C(O)NHS(O)₂alkyl.In other embodiments, R^(b) is hydrogen, CO₂R^(x), CH₂CO₂R^(x),tetrazole, or oxadiazolone. In further embodiments, R^(b) is hydrogen,CO₂H, CH₂CO₂H, tetrazole, or 1,2,4-oxadiazol-5(4H)-one. In furtherembodiments, R^(b) is hydrogen.

In some embodiments of Formula (Ib), R^(c) is H, C₁-C₆ alkyl, C₁-C₆haloalkyl, halogen, —CN, —OR^(x), —CO₂R^(x), or NO₂. In otherembodiments, R^(c) is C₁-C₆ alkyl, C₁-C₆ haloalkyl, halogen, —CN,—OR^(x), —CO₂R^(x), or NO₂. In other embodiments, R^(c) is halogen, —CN,—OR^(x), or C₁-C₆ alkyl. In other embodiments, R^(c) is halogen, —CN,—OR^(x), or C₁-C₃ alkyl. In other embodiments, R^(c) is H, —CN, orhalogen. In other embodiments, R^(c) is —CN or halogen.

In some embodiments of Formula (Ib), R^(d) is methyl, optionallysubstituted 5- to 10-membered aryl, optionally substituted 5- or6-membered heteroaryl, or optionally substituted 5- or 6-memberedcarbocycle. In other embodiments, R^(d) is methyl, optionallycyclohexyl, optionally substituted pyridinyl, optionally substitutedthiazolyl, optionally substituted phenyl, or optionally substitutedthienyl. In other embodiments, R^(d) is cyclohexyl, pyridinyl,thiazolyl, phenyl, or thienyl, wherein each is optionally substitutedwith one or more substituents independently selected from halogen, C₁-C₆alkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, C₁-C₆haloalkoxy, —OH, CN, and amino. In other embodiments, R^(d) iscyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each isoptionally substituted with one or more substituents independentlyselected from halogen, C₁-C₆ alkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy,C₁-C₆ haloalkyl, and C₁-C₆ haloalkoxy. In other embodiments, R^(d) iscyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each isoptionally substituted with one or more halogen. In other embodiments,R^(d) is methyl, cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl.In yet other embodiments, R^(d) is cyclohexyl, pyridinyl, thiazolyl,phenyl, or thienyl. In other embodiments, R^(d) is cyclohexyl,pyridinyl, thiazolyl, phenyl, 4-chlorophenyl, 4-methylphenyl, orthienyl.

In some embodiments of Formula (Ib), each R^(e) is independently C₁-C₆alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, halogen, —OR^(y), C₁-C₆ haloalkyl,—NHR^(z), —OH, or —CN. In other embodiments, C₁-C₄ alkyl, C₂-C₄ alkenyl,C₂-C₄ alkynyl, halogen, —OR^(y), C₁-C₄ haloalkyl, —NHR^(z), —OH, or —CN.

In some embodiments of Formula (Ib), R^(x) is hydrogen or C₁-C₆ alkyl.In other embodiments, R^(x) is hydrogen or C₁-C₃ alkyl. In furtherembodiments, R^(x) is hydrogen, methyl, ethyl, n-propyl, or iso-propyl.

In some embodiments of Formula (Ib), R^(y) is independently hydrogen,C₁-C₆ alkyl, or C₁-C₆ haloalkyl. In other embodiments, R^(y) ishydrogen, C₁-C₃ alkyl, or C₁-C₃ haloalkyl.

In some embodiments of Formula (Ib), each R^(z) is independentlyhydrogen, C₁-C₆ alkyl, or C₁-C₆ haloalkyl. In other embodiments, R^(z)is hydrogen, C₁-C₃ alkyl, or C₁-C₃ haloalkyl.

In some embodiments of Formula (Ib), n is 0, 1, 2, or 3. In otherembodiments, n is 0 or 1. In further embodiments, n is 0.

In some embodiments of Formula (Ib), one of R^(a) and R^(b) is hydrogenand the other is CO₂R^(x), CH₂CO₂R^(x), tetrazole, or oxadiazolone. Inother embodiments, R^(b) is hydrogen and R^(a) is CH₂CO₂H, tetrazole, or(1,2,4-oxadiazol-5(4H)-one).

In some embodiments of Formula (Ib), R^(b) is hydrogen, R^(c) is —CN,R^(d) is thienyl, and R^(a) is CH₂CO₂H, tetrazole, or(1,2,4-oxadiazol-5(4H)-one).

In some embodiments of Formula (Ib), R^(c) is halogen, R^(a) is —CO₂H,and R^(b) is H. In other embodiments, R^(c) is —Br, R^(a) is —CO₂H, andR^(b) is H. In further embodiments, R^(c) is —Cl, R^(a) is —CO₂H, andR^(b) is H.

In some embodiments of Formula (Ib), R^(c) is halogen, R^(a) istetrazole, and R^(b) is H. In other embodiments, R^(c) is —Br, R^(a) istetrazole, and R^(b) is H. In further embodiments, R^(c) is —Cl, R^(a)is tetrazole, and R^(b) is H.

In some embodiments of Formula (Ib), R^(c) is halogen, R^(a) is—CH₂CO₂H, and R^(b) is H. In other embodiments, R^(c) is —Br, R^(a) is—CH₂CO₂H, and R^(b) is H. In further embodiments, R^(c) is —Cl, R^(a) is—CH₂CO₂H, and R^(b) is H.

In some embodiments of Formula (Ib), R^(c) is halogen, R^(a) is(1,2,4-oxadiazol-5(4H)-one), and R^(b) is H. In other embodiments, R^(c)is —Br, R^(a) is (1,2,4-oxadiazol-5(4H)-one), and R^(b) is H. In otherembodiments, R^(c) is —Cl, R^(a) is (1,2,4-oxadiazol-5(4H)-one), andR^(b) is H.

In some embodiments of Formula (Ib), R^(c) is —CN, R^(a) is —CO₂H, andR^(b) is H. In other embodiments, R^(c) is —CN, R^(a) is —CH₂CO₂H, andR^(b) is H. In other embodiments, R^(c) is —CN, R^(a) is tetrazole, andR^(b) is H. In yet other embodiments, R^(c) is —CN, R^(a) is(1,2,4-oxadiazol-5(4H)-one), and R^(b) is H.

In some embodiments of Formula (Ib), R^(c) is not hydrogen or —CN andR^(d) is optionally substituted phenyl. In other embodiments, R^(c) isnot C₁-C₆ alkyl and R^(d) is methyl. In other embodiments, R^(c) is not—CN and R^(d) is 2-furyl.

In some embodiments of Formula (Ib), R^(c) is not hydrogen or —CN whenand R^(d) is optionally substituted phenyl.

In some embodiments of Formula (Ib), R^(c) is not C₁-C₆ alkyl when R^(d)is methyl.

In some embodiments of Formula (Ib), R^(c) is not —CN when R^(d) is2-furyl.

In another embodiment, the compound of Formula (I) is represented byFormula (II):

or a pharmaceutically acceptable salt thereof,

wherein:

R^(c) is halogen, —CN, —OR^(y), or C₁-C₆ alkyl;

R^(d) is methyl, optionally substituted 5- to 10-membered aryl,optionally substituted 5- or 6-membered heteroaryl, or optionallysubstituted 5- or 6-membered carbocycle; and

R^(x) is hydrogen or C₁-C₆ alkyl

with the proviso that R^(c) is not —CN when and R^(d) is optionallysubstituted phenyl, R^(c) is not C₁-C₆ alkyl when R^(d) is methyl, andthat R^(c) is not —CN when R^(d) is 2-furyl.

In some embodiments of Formula (II), R^(c) is halogen, —CN, —OR^(y), orC₁-C₆ alkyl; R^(d) is methyl, optionally substituted 5- to 10-memberedaryl, optionally substituted 5- or 6-membered heteroaryl, or optionallysubstituted 5- or 6-membered carbocycle; and R^(x) is hydrogen or C₁-C₆alkyl with the proviso that R^(c) is not C₁-C₆ alkyl when R^(d) ismethyl, and that R^(c) is not —CN when R^(d) is 2-furyl.

In some embodiments of Formula (II), R^(c) is halogen, —CN, —OR^(x), orC₁-C₆ alkyl. In other embodiments, R^(c) is halogen, —CN, —OR^(x), orC₁-C₃ alkyl. In further embodiments, R^(c) is —CN or halogen.

In some embodiments of Formula (II), R^(d) is methyl, optionallysubstituted 5- to 10-membered aryl, optionally substituted 5- or6-membered heteroaryl, or optionally substituted 5- or 6-memberedcarbocycle. In other embodiments, R^(d) is cyclohexyl, pyridinyl,thiazolyl, phenyl, or thienyl, wherein each is optionally substitutedwith one or more substituents independently selected from halogen, C₁-C₆alkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, C₁-C₆haloalkoxy, —OH, CN, and amino. In other embodiments, R^(d) iscyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each isoptionally substituted with one or more substituents independentlyselected from halogen, C₁-C₆ alkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy,C₁-C₆ haloalkyl, and C₁-C₆ haloalkoxy. In other embodiments, R^(d) iscyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each isoptionally substituted with one or more halogen. In further embodiments,R^(d) is methyl, cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl.

In some embodiments of Formula (II), R^(b) is hydrogen, CO₂R^(x),CH₂CO₂R^(x), tetrazole, or oxadiazolone. In other embodiments, R^(b) ishydrogen, CO₂H, CH₂CO₂H, tetrazole, or 1,2,4-oxadiazol-5(4H)-one. Infurther embodiments, R^(b) is hydrogen.

In some embodiments of Formula (II), R^(a) is hydrogen, CO₂R^(x),CH₂CO₂R^(x), tetrazole, or oxadiazolone. In further embodiments, R^(a)is hydrogen, CO₂H, CH₂CO₂H, tetrazole, or 1,2,4-oxadiazol-5(4H)-one.

In some embodiments of Formula (II), each R^(e) is independently C₁-C₆alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, halogen, —OR^(y), C₁-C₆ haloalkyl,—NHR^(z), —OH, or —CN.

In some embodiments of Formula (II), each R^(y) is independentlyhydrogen, C₁-C₆ alkyl, or C₁-C₆ haloalkyl.

In some embodiments of Formula (II), each R^(z) is independentlyhydrogen, C₁-C₆ alkyl, or C₁-C₆ haloalkyl.

In some embodiments of Formula (II), n is 0, 1, 2, or 3. In otherembodiments, n is 0 or 1. In further embodiments, n is 0.

In some embodiments of Formula (II), one of R^(a) and R^(b) is hydrogenand the other is CO₂R^(x), CH₂CO₂R^(x), tetrazole, or oxadiazolone. Inother embodiments, R^(b) is hydrogen and R^(a) is CH₂CO₂H, tetrazole, or(1,2,4-oxadiazol-5(4H)-one).

In some embodiments of Formula (II), R^(b) is hydrogen, R^(c) is —CN,R^(d) is thienyl, and R^(a) is CH₂CO₂H, tetrazole, or(1,2,4-oxadiazol-5(4H)-one).

In some embodiments of Formula (II), R^(c) is halogen, R^(a) is —CO₂H,and R^(b) is H. In other embodiments, R^(c) is —Br, R^(a) is —CO₂H, andR^(b) is H. In further embodiments, R^(c) is —Cl, R^(a) is —CO₂H, andR^(b) is H.

In some embodiments of Formula (II), R^(c) is halogen, R^(a) istetrazole, and R^(b) is H. In other embodiments, R^(c) is —Br, R^(a) istetrazole, and R^(b) is H. In further embodiments, R^(c) is —Cl, R^(a)is tetrazole, and R^(b) is H.

In some embodiments of Formula (II), R^(c) is halogen, R^(a) is—CH₂CO₂H, and R^(b) is H. In other embodiments, R^(c) is —Br, R^(a) is—CH₂CO₂H, and R^(b) is H. In further embodiments, R^(c) is —Cl, R^(a) is—CH₂CO₂H, and R^(b) is H.

In some embodiments of Formula (II), R^(c) is halogen, R^(a) is(1,2,4-oxadiazol-5(4H)-one), and R^(b) is H. In other embodiments, R^(c)is —Br, R^(a) is (1,2,4-oxadiazol-5(4H)-one), and R^(b) is H. In otherembodiments, R^(c) is —Cl, R^(a) is (1,2,4-oxadiazol-5(4H)-one), andR^(b) is H.

In some embodiments of Formula (II), R^(c) is —CN, R^(a) is —CO₂H, andR^(b) is H. In other embodiments, R^(c) is —CN, R^(a) is —CH₂CO₂H, andR^(b) is H. In other embodiments, R^(c) is —CN, R^(a) is tetrazole, andR^(b) is H. In yet other embodiments, R^(c) is —CN, R^(a) is(1,2,4-oxadiazol-5(4H)-one), and R^(b) is H.

In another embodiment, the compound of Formula (I) is represented byFormula (III):

or a pharmaceutically acceptable salt thereof

wherein:

R^(a) and R^(b) is hydrogen and the other is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)tetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r) isoxazol-3-ol,—(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH, —(CH₂)_(r)C(O)NHCN, or—(CH₂)_(r)C(O)NHS(O)₂alkyl;

R^(c) is H, C₁-C₆ alkyl, C₁-C₆ haloalkyl, halogen, —CN, —OR^(x),—CO₂R^(x), or NO₂;

R^(d) is methyl, optionally substituted 5- to 10-membered aryl,optionally substituted 5- or 6-membered heteroaryl, or optionallysubstituted 5- or 6-membered carbocycle;

each R^(x) is independently at each occurrence hydrogen or C₁-C₆ alkyl;

each R^(e) is independently C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,halogen, —OR^(y), C₁-C₆ haloalkyl, —NHR^(z), —OH, or —CN;

each R^(y) and R^(z) is independently hydrogen, C₁-C₆ alkyl, or C₁-C₆haloalkyl; and

n is 0, 1, 2, or 3.

In some embodiments of Formula (III),

one of R^(a) and R^(b) is hydrogen and the other is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)tetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r) isoxazol-3-ol,—(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH, —(CH₂)_(r)C(O)NHCN, or—(CH₂)_(r) C(O)NHS(O)₂alkyl;

R^(c) is C₁-C₆ alkyl, C₁-C₆ haloalkyl, halogen, —CN, —OR^(x), —CO₂R^(x),or NO₂;

R^(d) is methyl, optionally substituted 5- to 10-membered aryl,optionally substituted 5- or 6-membered heteroaryl, or optionallysubstituted 5- or 6-membered carbocycle;

each R^(x) is independently at each occurrence hydrogen or C₁-C₆ alkyl;

each R^(e) is independently C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,halogen, —OR^(y), C₁-C₆ haloalkyl, —NHR^(z), —OH, or —CN.

In some embodiments of formula (III),

one of R^(a) and R^(b) is hydrogen and the other is CO₂R^(x),CH₂CO₂R^(x), tetrazole, or oxadiazolone;

R^(c) is halogen, —CN, —OR^(x), or C₁-C₆ alkyl;

R^(d) is methyl, optionally substituted 5- to 10-membered aryl,optionally substituted 5- or 6-membered heteroaryl, or optionallysubstituted 5- or 6-membered carbocycle; and

R^(x) is hydrogen or C₁-C₆ alkyl;

each R^(e) is independently C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,halogen, —OR^(y), C₁-C₆ haloalkyl, —NHR^(z), —OH, or —CN;

each R^(y) and R^(z) is independently hydrogen, C₁-C₆ alkyl, or C₁-C₆haloalkyl; and

n is 0, 1, 2, or 3;

with the proviso that R^(c) is not hydrogen or —CN when R^(d) isoptionally substituted phenyl and that R^(c) is not —CN when R^(d) is2-furyl.

In some embodiments of Formula (III), R^(a) is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)tetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r) isoxazol-3-ol,—(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH, —(CH₂)_(r)C(O)NHCN, or—(CH₂)_(r)C(O)NHS(O)₂alkyl. In other embodiments, R^(a) is—(CH₂)_(r)CO₂R^(x), —OCH₂CO₂R^(x), tetrazole, —(CH₂)tetrazole,oxadiazolone, —(CH₂)oxadiazolone, tetrazolone, —(CH₂)tetrazolone,thiadiazolol, —(CH₂)thiadiazolol, isoxazol-3-ol, —(CH₂) isoxazol-3-ol,—P(O)(OH)OR^(x), —(CH₂)P(O)(OH)OR^(x), —S(O)₂OH, —(CH₂)S(O)₂OH,—C(O)NHCN —(CH₂)C(O)NHCN, —C(O)NHS(O)₂alkyl, or —(CH₂)C(O)NHS(O)₂alkyl.In other embodiments, R^(a) is hydrogen, CO₂R^(x), CH₂CO₂R^(x),tetrazole, or oxadiazolone. In further embodiments, R^(a) is hydrogen,CO₂H, CH₂CO₂H, tetrazole, or 1,2,4-oxadiazol-5(4H)-one.

In some embodiments of Formula (III), R^(b) is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)tetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r) isoxazol-3-ol,—(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH, —(CH₂)_(r)C(O)NHCN, or—(CH₂)_(r)C(O)NHS(O)₂alkyl. In other embodiments, R^(b) is—(CH₂)_(r)CO₂R^(x), —OCH₂CO₂R^(x), tetrazole, —(CH₂)tetrazole,oxadiazolone, —(CH₂)oxadiazolone, tetrazolone, —(CH₂)tetrazolone,thiadiazolol, —(CH₂)thiadiazolol, isoxazol-3-ol, —(CH₂) isoxazol-3-ol,—P(O)(OH)OR^(x), —(CH₂)P(O)(OH)OR^(x), —S(O)₂OH, —(CH₂)S(O)₂OH,—C(O)NHCN —(CH₂)C(O)NHCN, —C(O)NHS(O)₂alkyl, or —(CH₂)C(O)NHS(O)₂alkyl.In other embodiments, R^(b) is hydrogen, CO₂R^(x), CH₂CO₂R^(x),tetrazole, or oxadiazolone. In further embodiments, R^(b) is hydrogen,CO₂H, CH₂CO₂H, tetrazole, or 1,2,4-oxadiazol-5(4H)-one. In furtherembodiments, R^(b) is hydrogen.

In some embodiments of Formula (III), R^(c) is H, C₁-C₆ alkyl, C₁-C₆haloalkyl, halogen, —CN, —OR^(x), —CO₂R^(x), or NO₂. In otherembodiments, R^(c) is C₁-C₆ alkyl, C₁-C₆ haloalkyl, halogen, —CN,—OR^(x), —CO₂R^(x), or NO₂. In other embodiments, R^(c) is halogen, —CN,—OR^(x), or C₁-C₆ alkyl. In other embodiments, R^(c) is halogen, —CN,—OR^(x), or C₁-C₃ alkyl. In other embodiments, R^(c) is H, —CN, orhalogen. In other embodiments, R^(c) is —CN or halogen.

In some embodiments of Formula (III), R^(d) is methyl, optionallysubstituted 5- to 10-membered aryl, optionally substituted 5- or6-membered heteroaryl, or optionally substituted 5- or 6-memberedcarbocycle. In other embodiments, R^(d) is methyl, optionallycyclohexyl, optionally substituted pyridinyl, optionally substitutedthiazolyl, optionally substituted phenyl, or optionally substitutedthienyl. In other embodiments, R^(d) is cyclohexyl, pyridinyl,thiazolyl, phenyl, or thienyl, wherein each is optionally substitutedwith one or more substituents independently selected from halogen, C₁-C₆alkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, C₁-C₆haloalkoxy, —OH, CN, and amino. In other embodiments, R^(d) iscyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each isoptionally substituted with one or more substituents independentlyselected from halogen, C₁-C₆ alkyl, C₁-C₆ hydroxyalkyl, C₁-C₆ alkoxy,C₁-C₆ haloalkyl, and C₁-C₆ haloalkoxy. In other embodiments, R^(d) iscyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl, wherein each isoptionally substituted with one or more halogen. In other embodiments,R^(d) is methyl, cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl.In yet other embodiments, R^(d) is cyclohexyl, pyridinyl, thiazolyl,phenyl, or thienyl. In other embodiments, R^(d) is cyclohexyl,pyridinyl, thiazolyl, phenyl, 4-chlorophenyl, 4-methylphenyl, orthienyl.

In some embodiments of Formula (III), each R^(e) is independently C₁-C₆alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, halogen, —OR^(y), C₁-C₆ haloalkyl,—NHR^(z), —OH, or —CN. In other embodiments, C₁-C₄ alkyl, C₂-C₄ alkenyl,C₂-C₄ alkynyl, halogen, —OR^(y), C₁-C₄ haloalkyl, —NHR^(z), —OH, or —CN.

In some embodiments of Formula (III), R^(x) is hydrogen or C₁-C₆ alkyl.In other embodiments, R^(x) is hydrogen or C₁-C₃ alkyl. In furtherembodiments, R^(x) is hydrogen, methyl, ethyl, n-propyl, or iso-propyl.

In some embodiments of Formula (III), R^(y) is independently hydrogen,C₁-C₆ alkyl, or C₁-C₆ haloalkyl. In other embodiments, R^(y) ishydrogen, C₁-C₃ alkyl, or C₁-C₃ haloalkyl.

In some embodiments of Formula (III), each R^(z) is independentlyhydrogen, C₁-C₆ alkyl, or C₁-C₆ haloalkyl. In other embodiments, R^(z)is hydrogen, C₁-C₃ alkyl, or C₁-C₃ haloalkyl.

In some embodiments of Formula (III), n is 0, 1, 2, or 3. In otherembodiments, n is 0 or 1. In further embodiments, n is 0.

In some embodiments of Formula (III), one of R^(a) and R^(b) is hydrogenand the other is CO₂R^(x), CH₂CO₂R^(x), tetrazole, or oxadiazolone. Inother embodiments, R^(b) is hydrogen and R^(a) is CH₂CO₂H, tetrazole, or(1,2,4-oxadiazol-5(4H)-one).

In some embodiments of Formula (III), R^(b) is hydrogen, R^(c) is —CN,R^(d) is thienyl, and R^(a) is CH₂CO₂H, tetrazole, or(1,2,4-oxadiazol-5(4H)-one).

In some embodiments of Formula (III), R^(c) is halogen, R^(a) is —CO₂H,and R^(b) is H. In other embodiments, R^(c) is —Br, R^(a) is —CO₂H, andR^(b) is H. In further embodiments, R^(c) is —Cl, R^(a) is —CO₂H, andR^(b) is H.

In some embodiments of Formula (III), R^(c) is halogen, R^(a) istetrazole, and R^(b) is H. In other embodiments, R^(c) is —Br, R^(a) istetrazole, and R^(b) is H. In further embodiments, R^(c) is —Cl, R^(a)is tetrazole, and R^(b) is H.

In some embodiments of Formula (III), R^(c) is halogen, R^(a) is—CH₂CO₂H, and R^(b) is H. In other embodiments, R^(c) is —Br, R^(a) is—CH₂CO₂H, and R^(b) is H. In further embodiments, R^(c) is —Cl, R^(a) is—CH₂CO₂H, and R^(b) is H.

In some embodiments of Formula (III), R^(c) is halogen, R^(a) is(1,2,4-oxadiazol-5(4H)-one), and R^(b) is H. In other embodiments, R^(c)is —Br, R^(a) is (1,2,4-oxadiazol-5(4H)-one), and R^(b) is H. In otherembodiments, R^(c) is —Cl, R^(a) is (1,2,4-oxadiazol-5(4H)-one), andR^(b) is H.

In some embodiments of Formula (III), R^(c) is —CN, R^(a) is —CO₂H, andR^(b) is H. In other embodiments, R^(c) is —CN, R^(a) is —CH₂CO₂H, andR^(b) is H. In other embodiments, R^(c) is —CN, R^(a) is tetrazole, andR^(b) is H. In yet other embodiments, R^(c) is —CN, R^(a) is(1,2,4-oxadiazol-5(4H)-one), and R^(b) is H.

In some embodiments of Formula (III), R^(c) is not hydrogen or —CN whenR^(d) is optionally substituted phenyl, R^(c) is not C₁-C₆ alkyl whenR^(d) is methyl, and that R^(c) is not —CN when R^(d) is 2-furyl.

In some embodiments of Formula (I), (Ia), (Ib), (II) and (III), one ofR^(a) or R^(b) is a carboxylic acid or a carboxylic acid bioisostere.

In some embodiments of Formula (I), (Ia), (Ib), (II) and (III), R^(a) is—CO₂H, —(CH₂)CO₂H, or —OCH₂CO₂H. In other embodiments, R^(a) is —CO₂CH₃,—CO₂CH₂CH₃, —CO₂CH₂CH₂CH₃, —CO₂CH(CH₃)₂, —(CH₂)CO₂CH₃, —(CH₂)CO₂CH₂CH₃,—(CH₂)CO₂CH₂CH₂CH₃, or —(CH₂)CO₂CH(CH₃)₂.

In some embodiments of Formula (I), (Ia), (Ib), (II) and (III), R^(a) is—P(O)(OH)OH, —(CH₂)P(O)(OH)OH, —P(O)(OH)OCH₃, —P(O)(OH)OCH₂CH₃,—P(O)(OH)OCH₂CH₂CH₃, —P(O)(OH)OCH(CH₃)₂, —(CH₂) P(O)(OH)OCH₃,—(CH₂)P(O)(OH)OCH₂CH₃, —(CH₂)P(O)(OH)OCH₂CH₂CH₃, or—(CH₂)P(O)(OH)OCH(CH₃)₂.

In some embodiments of Formula (I), (Ia), (Ib), (II) and (III), R^(a) is—S(O)₂OH, —(CH₂)S(O)₂OH, —C(O)NHCN, or —(CH₂)C(O)NHCN.

In some embodiments of Formula (I), (Ia), (Ib), (II) and (III), R^(a) is—C(O)NHS(O)₂CH₃, —C(O)NHS(O)₂CH₂CH₃, —C(O)NHS(O)₂CH₂CH₂CH₃,—C(O)NHS(O)₂CH(CH₃)₂, —(CH₂)C(O)NHS(O)₂CH₃, —(CH₂)C(O)NHS(O)₂CH₂CH₃,—(CH₂)C(O)NHS(O)₂CH₂CH₂CH₃, or —(CH₂)C(O)NHS(O)₂CH(CH₃)₂.

In some embodiments of Formula (I), (Ia), (Ib), (II) and (III), R^(a) is

-   -   In some embodiments of Formula (I), (Ia), (Ib), (II), and (III),        R^(a) is

In some embodiments of Formula (I), (Ia), (Ib), (II) and (III), R^(b) is—CO₂H, —(CH₂)CO₂H, or —OCH₂CO₂H. In other embodiments, R^(b) is —CO₂CH₃,—CO₂CH₂CH₃, —CO₂CH₂CH₂CH₃, —CO₂CH(CH₃)₂, —(CH₂)CO₂CH₃, —(CH₂)CO₂CH₂CH₃,—(CH₂)CO₂CH₂CH₂CH₃, or —(CH₂)CO₂CH(CH₃)₂.

In some embodiments of Formula (I), (Ia), (Ib), (II) and (III), R^(b) is—P(O)(OH)OH, —(CH₂)P(O)(OH)OH, —P(O)(OH)OCH₃, —P(O)(OH)OCH₂CH₃,—P(O)(OH)OCH₂CH₂CH₃, —P(O)(OH)OCH(CH₃)₂, —(CH₂) P(O)(OH)OCH₃,—(CH₂)P(O)(OH)OCH₂CH₃, —(CH₂)P(O)(OH)OCH₂CH₂CH₃, or—(CH₂)P(O)(OH)OCH(CH₃)₂.

In some embodiments of Formula (I), (Ia), (Ib), (II) and (III), R^(b) is—S(O)₂OH, —(CH₂)S(O)₂OH, —C(O)NHCN, or —(CH₂)C(O)NHCN.

In some embodiments of Formula (I), (Ia), (Ib), (II) and (III), R^(b) is—C(O)NHS(O)₂CH₃, —C(O)NHS(O)₂CH₂CH₃, —C(O)NHS(O)₂CH₂CH₂CH₃,—C(O)NHS(O)₂CH(CH₃)₂, —(CH₂)C(O)NHS(O)₂CH₃, —(CH₂)C(O)NHS(O)₂CH₂CH₃,—(CH₂)C(O)NHS(O)₂CH₂CH₂CH₃, or —(CH₂)C(O)NHS(O)₂CH(CH₃)₂.

In some embodiments of Formula (I), (Ia), (Ib), (II) and (III), R^(b) is

In some embodiments of Formula (I), (Ia), (Ib), (II), and (III), R^(b)is

In some embodiments, the compound of Formula (I) is a compound havingany one of the following Formulae:

or a pharmaceutically acceptable salt thereof.

The above definition of the compounds of Formula (I) is referred toherein by the expressions “compound of Formula (I)” as defined herein,or simply “compounds of Formula (I)”, etc. The above definition of thecompounds of Formula (Ia) is referred to herein by the expressions“compound of Formula (Ia)” as defined herein, or simply “compounds ofFormula (Ia)”, etc. The above definition of the compounds of Formula(Ib) is referred to herein by the expressions “compound of Formula (Ib)”as defined herein, or simply “compounds of Formula (Ib)”, etc. The abovedefinition of the compounds of Formula (II) is referred to herein by theexpressions “compound of Formula (II)” as defined herein, or simply“compounds of Formula (II)”, etc. The above definition of the compoundsof Formula (III) is referred to herein by the expressions “compound ofFormula (III)” as defined herein, or simply “compounds of Formula(III)”, etc. It should be understood, that such references are intendedto encompass not only the above general formula, but also each and everyof the embodiments, etc. discussed in the following. It should also beunderstood, that unless stated to the opposite, such references alsoencompass isomers, mixtures of isomers, pharmaceutically acceptablesalts, solvates and prodrugs of the compounds of Formula (I), Formula(Ia), Formula (Ib), Formula (II), and Formula (III).

Definitions

The term “alkyl” as used herein refers to a saturated, straight orbranched hydrocarbon chain. The hydrocarbon chain preferably containsfrom one to eight carbon atoms (C₁₋₈-alkyl), more preferred from one tosix carbon atoms (C₁₋₆-alkyl), in particular from one to four carbonatoms (C₁₋₄-alkyl), including methyl, ethyl, propyl, isopropyl, butyl,isobutyl, secondary butyl, tertiary butyl, pentyl, isopentyl, neopentyl,tertiary pentyl, hexyl, isohexyl, heptyl and octyl. In a preferredembodiment “alkyl” represents a C₁₋₄-alkyl group, which may inparticular include methyl, ethyl, propyl, isopropyl, butyl, isobutyl,secondary butyl, and tertiary butyl. Correspondingly, the term“alkylene” means the corresponding biradical (-alkyl-).

The term “cycloalkyl” or “carbocycle” as used herein refers to a cyclicalkyl group, preferably containing from three to ten carbon atoms(C₃₋₁₀-cycloalkyl or C₃₋₁₀-carbocycle), such as from three to eightcarbon atoms (C₃₋₈-cycloalkyl or C₃₋₁₀-carbocycle), preferably fromthree to six carbon atoms (C₃₋₆-cycloalkyl or C₃₋₁₀-carbocycle),including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyland cyclooctyl. Furthermore, the term “cycloalkyl” as used herein mayalso include polycyclic groups such as for example bicyclo[2.2.2]octyl,bicyclo[2.2.1]heptanyl, decalinyl and adamantyl. Correspondingly, theterm “cycloalkylene” means the corresponding biradical (-cycloalkyl-).Alkyl and cycloalkyl groups may be optionally substituted with 1-4substituents. Examples of substituents on alkyl groups include, but arenot limited to, alkyl, alkenyl, alkynyl, halogen, haloalkyl, alkoxy,heteroaryl, aryl, carbocyclyl, hydroxyl, carbamoyl, oxo, and —CN.

The term “alkenyl” as used herein refers to a straight or branchedhydrocarbon chain or cyclic hydrocarbons containing one or more doublebonds, including di-enes, tri-enes and poly-enes. Typically, the alkenylgroup comprises from two to eight carbon atoms (C₂₋₈-alkenyl), such asfrom two to six carbon atoms (C₂₋₆-alkenyl), in particular from two tofour carbon atoms (C₂₋₄-alkenyl), including at least one double bond.Examples of alkenyl groups include ethenyl; 1- or 2-propenyl; 1-, 2- or3-butenyl, or 1,3-but-dienyl; 1-, 2-, 3-, 4- or 5-hexenyl, or1,3-hex-dienyl, or 1,3,5-hex-trienyl; 1-, 2-, 3-, 4-, 5-, 6-, or7-octenyl, or 1,3-octadienyl, or 1,3,5-octatrienyl, or1,3,5,7-octatetraenyl, or cyclohexenyl. Correspondingly, the term“alkenylene” means the corresponding biradical (-alkenyl-). Alkenylgroups may be optionally substituted with 1-4 substituents. Examples ofsubstituents on alkenyl groups include, but are not limited to, alkyl,alkenyl, alkynyl, halogen, haloalkyl, alkoxy, heteroaryl, aryl,carbocyclyl, hydroxyl, carbamoyl, oxo, and —CN.

The term “alkynyl” as used herein refers to a straight or branchedhydrocarbon chain containing one or more triple bonds, includingdi-ynes, tri-ynes and poly-ynes. Typically, the alkynyl group comprisesof from two to eight carbon atoms (C₂₋₈-alkynyl), such as from two tosix carbon atoms (C₂₋₆-alkynyl), in particular from two to four carbonatoms (C₂₋₄-alkynyl), including at least one triple bond. Examples ofpreferred alkynyl groups include ethynyl; 1- or 2-propynyl; 1-, 2- or3-butynyl, or 1,3-but-diynyl; 1-, 2-, 3-, 4- or 5-hexynyl, or1,3-hex-diynyl, or 1,3,5-hex-triynyl; 1-, 2-, 3-, 4-, 5-, 6-, or7-octynyl, or 1,3-oct-diynyl, or 1,3,5-oct-triynyl, or1,3,5,7-oct-tetraynyl. Correspondingly, the term “alkynylene” means thecorresponding biradical (-alkynyl-). Alkynyl groups may be optionallysubstituted with 1-4 substituents. Examples of substituents on alkynylgroups include, but are not limited to, alkyl, alkenyl, alkynyl,halogen, haloalkyl, alkoxy, heteroaryl, aryl, carbocyclyl, hydroxyl,carbamoyl, oxo, and —CN.

The terms “halo” and “halogen” as used herein refer to fluoro, chloro,bromo or iodo. Thus a trihalomethyl group represents, e.g., atrifluoromethyl group, or a trichloromethyl group. Preferably, the terms“halo” and “halogen” designate fluoro or chloro.

The term “haloalkyl” as used herein refers to an alkyl group, as definedherein, which is substituted one or more times with one or more halogen.Examples of haloalkyl groups include, but are not limited to,trifluoromethyl, difluoromethyl, pentafluoroethyl, trichloromethyl, etc.

The term “alkoxy” as used herein refers to an “alkyl-O—” group, whereinalkyl is as defined above.

The term “hydroxyalkyl” as used herein refers to an alkyl group (asdefined hereinabove), which alkyl group is substituted one or more timeswith hydroxy. Examples of hydroxyalkyl groups include HO—CH₂—,HO—CH₂—CH₂— and CH₃—CH(OH)—.

The term “oxy” as used herein refers to an “—O—” group.

The term “oxo” as used herein refers to an “═O” group.

The term “amine” as used herein refers to primary (R—NH₂, R≠H),secondary ((R)₂—NH, (R)₂≠H) and tertiary ((R)₃—N, R≠H) amines Asubstituted amine is intended to mean an amine where at least one of thehydrogen atoms has been replaced by the substituent.

The term “carbamoyl” as used herein refers to a “H₂N(═O)—” group.

The term “aryl”, as used herein, unless otherwise indicated, includescarbocyclic aromatic ring systems derived from an aromatic hydrocarbonby removal of a hydrogen atom. Aryl furthermore includes bi-, tri-andpolycyclic ring systems. Examples of preferred aryl moieties includephenyl, naphthyl, indenyl, indanyl, fluorenyl, biphenyl, indenyl,naphthyl, anthracenyl, phenanthrenyl, pentalenyl, azulenyl, andbiphenylenyl. Preferred “aryl” is phenyl, naphthyl or indanyl, inparticular phenyl, unless otherwise stated. Any aryl used may beoptionally substituted. Correspondingly, the term “arylene” means thecorresponding biradical (-aryl-). Aryl groups may be optionallysubstituted with 1-4 substituents. Examples of substituents on arylgroups include, but are not limited to, alkyl, alkenyl, alkynyl,halogen, haloalkyl, alkoxy, heteroaryl, aryl, carbocyclyl, hydroxyl, and—CN.

The term “heteroaryl”, as used herein, refers to aromatic groupscontaining one or more heteroatoms selected from O, S, and N, preferablyfrom one to four heteroatoms, and more preferably from one to threeheteroatoms. Heteroaryl furthermore includes bi-, tri-and polycyclicgroups, wherein at least one ring of the group is aromatic, and at leastone of the rings contains a heteroatom selected from O, S, and N.Heteroaryl also include ring systems substituted with one or more oxomoieties. Examples of preferred heteroaryl moieties includeN-hydroxytetrazolyl, N-hydroxytriazolyl, N-hydroxyimidazolyl, furanyl,triazolyl, pyranyl, thiadiazinyl, benzothiophenyl,dihydro-benzo[b]thiophenyl, xanthenyl, isoindanyl, acridinyl,benzisoxazolyl, quinolinyl, isoquinolinyl, phteridinyl, azepinyl,diazepinyl, imidazolyl, thiazolyl, carbazolyl, pyridinyl, pyridazinyl,pyrimidinyl, pyrazolyl, pyrazinyl, tetrazolyl, furyl, thienyl,isoxazolyl, oxazolyl, isothiazolyl, pyrrolyl, indolyl, benzimidazolyl,benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl,triazinyl, isoindolyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl,benzofurazanyl, benzothiophenyl, benzotriazolyl, benzothiazolyl,benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl,dihydroquinolyl, tetrahydroquinolyl, dihydroisoquinolyl,tetrahydroisoquinolyl, benzofuryl, furopyridinyl, pyrolopyrimidinyl,azaindolyl, pyrazolinyl, 1,2,4-oxadiazol-5(4H)-one, and pyrazolidinyl.Non-limiting examples of partially hydrogenated derivatives are1,2,3,4-tetrahydronaphthyl, 1,4-dihydronaphthyl, and 1-octalin.Correspondingly, the term “heteroarylene” means the correspondingbiradical (-heteroaryl-). Heteroaryl groups may be optionallysubstituted with 1-4 substituents. Examples of substituents onheteroaryl groups include, but are not limited to, alkyl, alkenyl,alkynyl, halogen, haloalkyl, alkoxy, heteroaryl, aryl, carbocyclyl,hydroxyl, and —CN.

The term “heterocyclyl” as used herein, refers to cyclic non-aromaticgroups containing one or more heteroatoms selected from O, S, and N,preferably from one to four heteroatoms, and more preferably from one tothree heteroatoms. Heterocyclyl furthermore includes bi-, tri-andpolycyclic non-aromatic groups, and at least one of the rings contains aheteroatom selected from O, S, and N. Heterocyclyl also include ringsystems substituted with one or more oxo moieties. Examples ofheterocyclic groups are oxetane, pyrrolidinyl, pyrrolyl, 3H-pyrrolyl,oxolanyl, furanyl, thiolanyl, thiophenyl, pyrazolyl, pyrazolidinyl,imidazolyl, imidazolidinyl, 3H-pyrazolyl, 1,2-oxazolyl, 1,3-oxazolyl,1,2-thiazolyl, 1,3-thiazolyl, 1,2,5-oxadiazolyl, piperidinyl, pyridinyl,oxanyl, 2-H-pyranyl, 4-H-pyranyl, thianyl, 2H-thiopyranyl, pyridazinyl,1,2-diazinanyl, pyrimidinyl, 1,3-diazinanyl, pyrazinyl, piperazinyl,1,4-dioxinyl, 1,4-dioxanyl, 1,3-diazinanyl, 1,4-oxazinyl, morpholinyl,thiomorpholinyl, 1,4-oxathianyl, benzofuranyl, isobenzofuranyl,indazolyl, benzimidazolyl, quinolinyl, isoquinolinyl, chromayl,isochromanyl, 4H-chromenyl, 1H-isochromenyl, cinnolinyl, quinazolinyl,quinoxalinyl, phthalazinyl, purinyl, naphthyridinyl, pteridinyl,indolizinyl, 1H-pyrrolizinyl, 4H-quinolizinyl andaza-8-bicyclo[3.2.1]octane. Correspondingly, the term “heterocyclylene”means the corresponding biradical (-heterocyclyl-). Heterocyclyl groupsmay be optionally substituted with 1-4 substituents. Examples ofsubstituents on heterocyclyl groups include, but are not limited, toalkyl, alkenyl, alkynyl, halogen, haloalkyl, alkoxy, heteroaryl, aryl,carbocyclyl, hydroxyl, and —CN.

The term “N-heterocyclic ring” as used herein, refers to a heterocyclylor a heteroaryl, as defined hereinabove, having at least one nitrogenatom, and being bound via a nitrogen atom. Examples of suchN-heterocyclic rings are pyrrolidinyl, pyrrolyl, 3H-pyrrolyl, pyrazolyl,pyrazolidinyl, imidazolyl, imidazolidinyl, 3H-pyrazolyl, 1,2-oxazolyl,1,2-thiazolyl, 1,3-thiazolyl, piperidinyl, pyridinyl, pyridazinyl,pyrazinyl, piperazinyl, morpholinyl, pyridinyl, pyridazinyl,pyrimidinyl, pyrazolyl, pyrazinyl, tetrazolyl, etc.

Isomers

In the present specification, the structural formula of the compoundrepresents a certain isomer for convenience in some cases, but thepresent disclosure includes all isomers, such as geometrical isomers,optical isomers based on an asymmetrical carbon, stereoisomers,tautomers, and the like. Accordingly, it should be understood that thedefinition of compounds of Formulae (I), (Ia), (Ib), (II) and (III)include each and every individual isomer corresponding to the Formula:Formulae (I), (Ia), (Ib), (II) and (III), including cis-trans isomers,stereoisomers and tautomers, as well as racemic mixtures of these andpharmaceutically acceptable salts thereof. Hence, the definition ofcompounds of Formulae (I), (Ia), (Ib), (II) and (III) are also intendedto encompass all R-and S-isomers of a chemical structure in any ratio,e.g., with enrichment (i.e., enantiomeric excess or diastereomericexcess) of one of the possible isomers and corresponding smaller ratiosof other isomers. In addition, a crystal polymorphism may be present forthe compounds represented by Formulae (I), (Ia), (Ib), (II) and (III).It is noted that any crystal form, crystal form mixture, or anhydride orhydrate thereof is included in the scope of the present disclosure.Furthermore, so-called metabolite which is produced by degradation ofthe present compound in vivo is included in the scope of the presentdisclosure.

“Isomerism” means compounds that have identical molecular formulae butdiffer in the sequence of bonding of their atoms or in the arrangementof their atoms in space. Isomers that differ in the arrangement of theiratoms in space are termed “stereoisomers”. Stereoisomers that are notmirror images of one another are termed “diastereoisomers”, andstereoisomers that are non-superimposable mirror images of each otherare termed “enantiomers” or sometimes optical isomers. A mixturecontaining equal amounts of individual enantiomeric forms of oppositechirality is termed a “racemic mixture”.

A carbon atom bonded to four non-identical substituents is termed a“chiral center”.

Chiral isomer” means a compound with at least one chiral center.Compounds with more than one chiral center may exist either as anindividual diastereomer or as a mixture of diastereomers, termed“diastereomeric mixture”. When one chiral center is present, astereoisomer may be characterized by the absolute configuration (R or S)of that chiral center. Absolute configuration refers to the arrangementin space of the substituents attached to the chiral center. Thesubstituents attached to the chiral center under consideration areranked in accordance with the Sequence Rule of Cahn, Ingold and Prelog.(Cahn et al., Angew. Chem. Inter. Edit. 1966, 5, 385; errata 511; Cahnet al., Angew. Chem. 1966, 78, 413; Cahn and Ingold, J. Chem. Soc. 1951(London), 612; Cahn et al., Experientia 1956, 12, 81; Cahn, J. Chem.Educ. 1964, 41, 116).

Diastereoisomers, i.e., non-superimposable stereochemical isomers, canbe separated by conventional means such as chromatography, distillation,crystallization or sublimation. The optical isomers can be obtained byresolution of the racemic mixtures according to conventional processes,for example by formation of diastereoisomeric salts by treatment with anoptically active acid or base. Examples of appropriate acids include,without limitation, tartaric, diacetyltartaric, dibenzoyltartaric,ditoluoyltartaric and camphorsulfonic acid. The mixture of diastereomerscan be separated by crystallization followed by liberation of theoptically active bases from these salts. An alternative process forseparation of optical isomers includes the use of a chiralchromatography column optimally chosen to maximize the separation of theenantiomers. Still another available method involves synthesis ofcovalent diastereoisomeric molecules by reacting compounds of Formula(I), (Ia), (Ib), (II) or (III) with an optically pure acid in anactivated form or an optically pure isocyanate. The synthesizeddiastereoisomers can be separated by conventional means such aschromatography, distillation, crystallization or sublimation, and thenhydrolyzed to obtain the enantiomerically pure compound. The opticallyactive compounds of Formulae (I), (Ia), (Ib), (II) and (III) canlikewise be obtained by utilizing optically active starting materialsand/or by utilizing a chiral catalyst. These isomers may be in the formof a free acid, a free base, an ester or a salt. Examples of chiralseparation techniques are given in Chiral Separation Techniques, APractical Approach, 2^(nd) ed. by G. Subramanian, Wiley-VCH, 2001.

“Geometric isomer” means the diastereomers that owe their existence tohindered rotation about double bonds. These configurations aredifferentiated in their names by the prefixes cis and trans, or Z and E,which indicate that the groups are on the same or opposite side of thedouble bond in the molecule according to the Cahn-Ingold-Prelog rules.

Furthermore, the structures and other compounds discussed in thisdisclosure include all atropic isomers thereof “Atropic isomers” are atype of stereoisomer in which the atoms of two isomers are arrangeddifferently in space. Atropic isomers owe their existence to arestricted rotation caused by hindrance of rotation of large groupsabout a central bond. Such atropic isomers typically exist as a mixture,however as a result of recent advances in chromatography techniques; ithas been possible to separate mixtures of two atropic isomers in selectcases.

“Tautomer” is one of two or more structural isomers that exist inequilibrium and is readily converted from one isomeric form to another.This conversion results in the formal migration of a hydrogen atomaccompanied by a switch of adjacent conjugated double bonds. Tautomersexist as a mixture of a tautomeric set in solution. In solid form,usually one tautomer predominates. In solutions where tautomerization ispossible, a chemical equilibrium of the tautomers will be reached. Theexact ratio of the tautomers depends on several factors, includingtemperature, solvent and pH. The concept of tautomers that areinterconvertable by tautomerizations is called tautomerism.

Of the various types of tautomerism that are possible, two are commonlyobserved. In keto-enol tautomerism a simultaneous shift of electrons anda hydrogen atom occurs. Ring-chain tautomerism arises as a result of thealdehyde group (—CHO) in a sugar chain molecule reacting with one of thehydroxy groups (—OH) in the same molecule to give it a cyclic(ring-shaped) form as exhibited by glucose.

Common tautomeric pairs are: ketone-enol, amide-nitrile, lactam-lactim,amide-imidic acid tautomerism in heterocyclic rings (e.g., innucleobases such as guanine, thymine and cytosine), amine-enamine andenamine-enamine. It is to be understood that the compounds of thepresent disclosure may be depicted as different tautomers. It shouldalso be understood that when compounds have tautomeric forms, alltautomeric forms are intended to be included in the scope of the presentdisclosure, and the naming of the compounds does not exclude anytautomer form.

The term “crystal polymorphs”, “polymorphs” or “crystal forms” meanscrystal structures in which a compound (or a salt or solvate thereof)can crystallize in different crystal packing arrangements, all of whichhave the same elemental composition. Different crystal forms usuallyhave different X-ray diffraction patterns, infrared spectral, meltingpoints, density hardness, crystal shape, optical and electricalproperties, stability and solubility. Recrystallization solvent, rate ofcrystallization, storage temperature, and other factors may cause onecrystal form to dominate. Crystal polymorphs of the compounds can beprepared by crystallization under different conditions.

Additionally, the compounds of the present disclosure, for example, thesalts of the compounds, can exist in either hydrated or unhydrated (theanhydrous) form or as solvates with other solvent molecules. Nonlimitingexamples of hydrates include monohydrates, dihydrates, etc. Nonlimitingexamples of solvates include ethanol solvates, acetone solvates, etc.

“Solvate” means solvent addition forms that contain eitherstoichiometric or non-stoichiometric amounts of solvent. Some compoundshave a tendency to trap a fixed molar ratio of solvent molecules in thecrystalline solid state, thus forming a solvate. If the solvent is waterthe solvate formed is a hydrate; and if the solvent is alcohol, thesolvate formed is an alcoholate. Hydrates are formed by the combinationof one or more molecules of water with one molecule of the substance inwhich the water retains its molecular state as H₂O.

The present disclosure is intended to include all isotopes of atomsoccurring in the present compounds. Isotopes include those atoms havingthe same atomic number but different mass numbers. By way of generalexample and without limitation, isotopes of hydrogen include tritium anddeuterium, and isotopes of carbon include C-13 and C-14.

Throughout the description and claims of this specification, the words“comprise” and “contain” and variations of the words, for example“comprising” and “comprises”, mean “including but not limited to” and donot exclude other moieties, additives, components, integers or steps.Throughout the description and claims of this specification, thesingular encompasses the plural unless the context otherwise requires.In particular, where the indefinite article is used, the specificationis to be understood as contemplating plurality as well as singularity,unless the context requires otherwise.

All references, including any patent or patent application, cited inthis specification are hereby incorporated by reference. No admission ismade that any reference constitutes prior art. Further, no admission ismade that any of the prior art constitutes part of the common generalknowledge in the art.

Method of Treatment

In another aspect, the present disclosure relates to a method oftreating a disease or disorder in whichα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD) plays arole comprising administering to the subject in need thereof atherapeutically effective amount of one or more compounds of Formula(I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof.

Another aspect of the present disclosure relates to a method of treatinga disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction comprising administering to the subject suffering from orsusceptible to developing a disease or disorder associated with ACMSDdysfunction a therapeutically effective amount of one or more compoundsof Formula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula(III), or a pharmaceutically acceptable salt thereof.

In another aspect, the present disclosure relates to a method ofpreventing a disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction comprising administering to the subject suffering from orsusceptible to developing a disease or disorder associated with ACMSDdysfunction a therapeutically effective amount of one or more compoundsof Formula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula(III), or a pharmaceutically acceptable salt thereof.

In yet another aspect, the present disclosure relates to a method ofreducing the risk of a disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction comprising administering to the subject suffering from orsusceptible to developing a disease or disorder associated with ACMSDdysfunction a therapeutically effective amount of one or more compoundsof Formula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula(III), or a pharmaceutically acceptable salt thereof.

Another aspect of the present disclosure relates to a method ofameliorating the risk of a disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction comprising administering to the subject suffering from orsusceptible to developing a disease or disorder associated with ACMSDdysfunction a therapeutically effective amount of one or more compoundsof Formula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula(III), or a pharmaceutically acceptable salt thereof.

In another aspect, the present disclosure relates to a method oftreating a disease or disorder in which nicotinamide adeninedinucleotide (NAD⁺) modulation plays a role comprising administering tothe subject in need thereof a therapeutically effective amount of one ormore compounds of Formula (I), Formula (Ia), Formula (Ib), Formula (II),or Formula (III), or a pharmaceutically acceptable salt thereof.

Another aspect of the present disclosure relates to a method ofpreventing a disease or disorder in which nicotinamide adeninedinucleotide (NAD⁺) modulation plays a role comprising administering tothe subject in need thereof a therapeutically effective amount of one ormore compounds of Formula (I), Formula (Ia), Formula (Ib), Formula (II),or Formula (III), or a pharmaceutically acceptable salt thereof.

In another aspect, the present disclosure relates to a method ofreducing the risk of a disease or disorder in which nicotinamide adeninedinucleotide (NAD⁺) modulation plays a role comprising administering tothe subject in need thereof a therapeutically effective amount of one ormore compounds of Formula (I), Formula (Ia), Formula (Ib), Formula (II),or Formula (III), or a pharmaceutically acceptable salt thereof.

Another aspect of the present disclosure relates to a method ofameliorating a disease or disorder in which nicotinamide adeninedinucleotide (NAD⁺) modulation plays a role comprising administering tothe subject in need thereof a therapeutically effective amount of one ormore compounds of Formula (I), Formula (Ia), Formula (Ib), Formula (II),or Formula (III), or a pharmaceutically acceptable salt thereof.

In another aspect, the present disclosure relates to a method oftreating a disease or disorder associated with reduced nicotinamideadenine dinucleotide (NAD⁺) levels comprising administering to thesubject suffering from or susceptible to developing a disease ordisorder associated with reduced NAD⁺ levels a therapeutically effectiveamount of one or more compounds of Formula (I), Formula (Ia), Formula(Ib), Formula (II), or Formula (III), or a pharmaceutically acceptablesalt thereof.

The present disclosure also relates to a method of preventing a diseaseor disorder associated with reduced nicotinamide adenine dinucleotide(NAD⁺) levels comprising administering to the subject suffering from orsusceptible to developing a disease or disorder associated with reducedNAD⁺ levels a therapeutically effective amount of one or more compoundsof Formula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula(III), or a pharmaceutically acceptable salt thereof.

Another aspect of the present disclosure relates to a method of reducingthe risk of a disease or disorder associated with reduced nicotinamideadenine dinucleotide (NAD⁺) levels comprising administering to thesubject suffering from or susceptible to developing a disease ordisorder associated with reduced NAD⁺ levels a therapeutically effectiveamount of one or more compounds of Formula (I), Formula (Ia), Formula(Ib), Formula (II), or Formula (III), or a pharmaceutically acceptablesalt thereof.

In another aspect, the present disclosure relates to a method ofameliorating a disease or disorder associated with reduced nicotinamideadenine dinucleotide (NAD⁺) levels comprising administering to thesubject suffering from or susceptible to developing a disease ordisorder associated with reduced NAD⁺ levels a therapeutically effectiveamount of one or more compounds of Formula (I), Formula (Ia), Formula(Ib), Formula (II), or Formula (III), or a pharmaceutically acceptablesalt thereof.

Another aspect of the present disclosure relates to a method of treatinga disorder associated with mitochondrial dysfunction comprisingadministering to the subject suffering from or susceptible to developinga metabolic disorder a therapeutically effective amount of one or morecompounds of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof. In oneembodiment, the disorder associated with mitochondrial dysfunction is aninherited mitochondrial disease, a common metabolic disorder, aneurodegenerative disease, an aging related disorder, a kidney disorder,or a chronic inflammatory disease. In a preferred embodiment, thedisorder associated with mitochondrial dysfunction is a common metabolicdisorder such as obesity or type II diabetes.

In another aspect, the present disclosure relates to a method ofpreventing a disorder associated with mitochondrial dysfunctioncomprising administering to the subject suffering from or susceptible todeveloping a metabolic disorder a therapeutically effective amount ofone or more compounds of Formula (I), Formula (Ia), Formula (Ib),Formula (II), or Formula (III), or a pharmaceutically acceptable saltthereof. In one embodiment, the disorder associated with mitochondrialdysfunction is an inherited mitochondrial disease, a common metabolicdisorder, a neurodegenerative disease, an aging related disorder, akidney disorder, or a chronic inflammatory disease. In a preferredembodiment, the disorder associated with mitochondrial dysfunction is acommon metabolic disorder such as obesity or type II diabetes.

Another aspect of the present disclosure relates to a method of reducingthe risk of a disorder associated with mitochondrial dysfunctioncomprising administering to the subject suffering from or susceptible todeveloping a metabolic disorder a therapeutically effective amount ofone or more compounds of Formula (I), Formula (Ia), Formula (Ib),Formula (II), or Formula (III), or a pharmaceutically acceptable saltthereof. In one embodiment, the disorder associated with mitochondrialdysfunction is an inherited mitochondrial disease, a common metabolicdisorder, a neurodegenerative disease, an aging related disorder, akidney disorder, or a chronic inflammatory disease. In a preferredembodiment, the disorder associated with mitochondrial dysfunction is acommon metabolic disorder such as obesity or type II diabetes.

Another aspect of the present disclosure relates to a method ofameliorating a disorder associated with mitochondrial dysfunctioncomprising administering to the subject suffering from or susceptible todeveloping a metabolic disorder a therapeutically effective amount ofone or more compounds of Formula (I), Formula (Ia), Formula (Ib),Formula (II), or Formula (III), or a pharmaceutically acceptable saltthereof. In one embodiment, the disorder associated with mitochondrialdysfunction is an inherited mitochondrial disease, a common metabolicdisorder, a neurodegenerative disease, an aging related disorder, akidney disorder, or a chronic inflammatory disease. In a preferredembodiment, the disorder associated with mitochondrial dysfunction is acommon metabolic disorder such as obesity or type II diabetes.

In another aspect, the present disclosure relates to a method ofpromoting oxidative metabolism comprising administering to the subjectsuffering from or susceptible to developing a metabolic disorder atherapeutically effective amount of one or more compounds of Formula(I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, that increases intracellularnicotinamide adenine dinucleotide (NAD⁺).

In yet another aspect, the present disclosure relates to a method forthe manufacture of a medicament for treating a disease or conditionmediated by ACMSD, wherein the medicament comprises a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof.

Another aspect of the present disclosure relates to a method for themanufacture of a medicament for preventing a disease or conditionmediated by ACMSD, wherein the medicament comprises a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof.

In another aspect, the present disclosure relates to a method for themanufacture of a medicament for reducing the risk of a disease orcondition mediated by ACMSD, wherein the medicament comprises a compoundof Formula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula(III), or a pharmaceutically acceptable salt thereof.

In yet another aspect, the present disclosure relates to a method forthe manufacture of a medicament for ameliorating a disease or conditionmediated by ACMSD, wherein the medicament comprises a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof

In another aspect, the present disclosure relates to a pharmaceuticalcomposition for use in a method for treating a disease or conditionmediated by ACMSD, wherein the medicament comprises a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof.

In another aspect, the present disclosure relates to a pharmaceuticalcomposition for use in a method for preventing a disease or conditionmediated by ACMSD, wherein the medicament comprises a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof.

In another aspect, the present disclosure relates to a pharmaceuticalcomposition for use in a method for reducing the risk of a disease orcondition mediated by ACMSD, wherein the medicament comprises a compoundof Formula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula(III), or a pharmaceutically acceptable salt thereof.

Another aspect of the present disclosure relates to a pharmaceuticalcomposition for use in a method for ameliorating a disease or conditionmediated by ACMSD, wherein the medicament comprises a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof.

In yet another aspect, the present disclosure relates to a compound foruse in a method for treating a disease or condition mediated by ACMSD,wherein the compound comprises a compound of Formula (I), Formula (Ia),Formula (Ib), Formula (II), or Formula (III), or a pharmaceuticallyacceptable salt thereof.

Another aspect of the present disclosure relates to a compound for usein a method for preventing a disease or condition mediated by ACMSD,wherein the compound comprises a compound of Formula (I), Formula (Ia),Formula (Ib), Formula (II), or Formula (III), or a pharmaceuticallyacceptable salt thereof.

In another aspect, the present disclosure relates to a compound for usein a method for reducing the risk of a disease or condition mediated byACMSD, wherein the compound comprises a compound of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof.

Another aspect of the present disclosure relates to a compound for usein a method for ameliorating a disease or condition mediated by ACMSD,wherein the compound comprises a compound of Formula (I), Formula (Ia),Formula (Ib), Formula (II), or Formula (III), or a pharmaceuticallyacceptable salt thereof.

Another aspect of the present disclosure relates to the use of acompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for treating, preventing or reducing therisk of a disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction.

In another aspect, the present disclosure relates to the use of acompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for treating the risk of a disease ordisorder associated with α-amino-β-carboxymuconate-ε-semialdehydedecarboxylase (ACMSD) dysfunction.

Another aspect of the present disclosure relates to the use of acompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for preventing a disease or disorderassociated with α-amino-β-carboxymuconate-ε-semialdehyde decarboxylase(ACMSD) dysfunction.

In another aspect, the present disclosure relates to the use of acompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for reducing the risk of a disease ordisorder associated with α-amino-β-carboxymuconate-ε-semialdehydedecarboxylase (ACMSD) dysfunction.

Another aspect of the present disclosure relates to the use of acompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for ameliorating a disease or disorderassociated with α-amino-β-carboxymuconate-ε-semialdehyde decarboxylase(ACMSD) dysfunction.

In another aspect, the present disclosure relates to the use of acompound of a compound of Formula (I), Formula (Ia), Formula (Ib),Formula (II), or Formula (III), or a pharmaceutically acceptable saltthereof in the manufacture of a medicament for treating, preventing orreducing the risk of a disease or disorder associated with reducednicotinamide adenine dinucleotide (NAD⁺) levels.

Another aspect of the present disclosure relates to the use of acompound of a compound of Formula (I), Formula (Ia), Formula (Ib),Formula (II), or Formula (III), or a pharmaceutically acceptable saltthereof in the manufacture of a medicament for treating a disease ordisorder associated with reduced nicotinamide adenine dinucleotide(NAD⁺) levels.

In another aspect, the present disclosure relates to the use of acompound of a compound of Formula (I), Formula (Ia), Formula (Ib),Formula (II), or Formula (III), or a pharmaceutically acceptable saltthereof in the manufacture of a medicament for preventing a disease ordisorder associated with reduced nicotinamide adenine dinucleotide(NAD⁺) levels.

Another aspect of the present disclosure relates to the use of acompound of a compound of Formula (I), Formula (Ia), Formula (Ib),Formula (II), or Formula (III), or a pharmaceutically acceptable saltthereof in the manufacture of a medicament reducing the risk of adisease or disorder associated with reduced nicotinamide adeninedinucleotide (NAD⁺) levels.

In another aspect, the present disclosure relates to the use of acompound of a compound of Formula (I), Formula (Ia), Formula (Ib),Formula (II), or Formula (III), or a pharmaceutically acceptable saltthereof in the manufacture of a medicament for ameliorating a disease ordisorder associated with reduced nicotinamide adenine dinucleotide(NAD⁺) levels.

Another aspect of the present disclosure relates to the use of acompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for treating, preventing or reducing therisk of a disorder associated with mitochondrial dysfunction.

In another aspect, the present disclosure relates to the use of acompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for treating a disorder associated withmitochondrial dysfunction.

Another aspect of the present disclosure relates to the use of acompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for preventing a disorder associated withmitochondrial dysfunction.

In another aspect, the present disclosure relates to the use of acompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for reducing the risk of a disorderassociated with mitochondrial dysfunction.

Another aspect of the present disclosure relates to the use of acompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof in themanufacture of a medicament for ameliorating a disorder associated withmitochondrial dysfunction.

In another aspect, the present disclosure relates to the use of acompound of a compound of Formula (I), Formula (Ia), Formula (Ib),Formula (II), or Formula (III), or a pharmaceutically acceptable saltthereof in the manufacture of a medicament for promoting oxidativemetabolism.

Another aspect of the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use in the manufactureof a medicament for treating, preventing or reducing the risk of adisease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction.

In another aspect, the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use in the manufactureof a medicament for treating a disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction.

Another aspect of the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use in the manufactureof a medicament for preventing a disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction.

In another aspect, the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use in the manufactureof a medicament for reducing the risk of a disease or disorderassociated with α-amino-β-carboxymuconate-ε-semialdehyde decarboxylase(ACMSD) dysfunction.

Another aspect of the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use in the manufactureof a medicament for ameliorating a disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction.

In another aspect, the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use as a medicamentfor treating, preventing or reducing the risk of a disease or disorderassociated with reduced nicotinamide adenine dinucleotide (NAD⁺) levels.

Another aspect of the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use as a medicamentfor treating a disease or disorder associated with reduced nicotinamideadenine dinucleotide (NAD⁺) levels.

In another aspect, the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use as a medicamentfor preventing a disease or disorder associated with reducednicotinamide adenine dinucleotide (NAD⁺) levels.

Another aspect of the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use as a medicamentfor reducing the risk of a disease or disorder associated with reducednicotinamide adenine dinucleotide (NAD⁺) levels.

In another aspect, the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use as a medicamentfor ameliorating a disease or disorder associated with reducednicotinamide adenine dinucleotide (NAD⁺) levels.

Another aspect of the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use as a medicamentfor treating, preventing or reducing the risk of a disorder associatedwith mitochondrial dysfunction.

In another aspect, the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use as a medicamentfor treating a disorder associated with mitochondrial dysfunction.

Another aspect of the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use as a medicamentfor preventing a disorder associated with mitochondrial dysfunction.

In another aspect, the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use as a medicamentfor reducing the risk of a disorder associated with mitochondrialdysfunction.

Another aspect of the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use as a medicamentfor ameliorating a disorder associated with mitochondrial dysfunction.

In another aspect, the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use as a medicamentfor promoting oxidative metabolism.

Another aspect of the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use in treating,preventing or reducing the risk of a disease or disorder associated withreduced nicotinamide adenine dinucleotide (NAD⁺) levels.

In another aspect, the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use in treating adisease or disorder associated with reduced nicotinamide adeninedinucleotide (NAD⁺) levels.

Another aspect of the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use in preventing adisease or disorder associated with reduced nicotinamide adeninedinucleotide (NAD⁺) levels.

In another aspect, the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use in reducing therisk of a disease or disorder associated with reduced nicotinamideadenine dinucleotide (NAD⁺) levels.

Another aspect of the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use in ameliorating adisease or disorder associated with reduced nicotinamide adeninedinucleotide (NAD⁺) levels.

In another aspect, the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use in for treating,preventing or reducing the risk of a disorder associated withmitochondrial dysfunction.

Another aspect of the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use in for treating adisorder associated with mitochondrial dysfunction.

In another aspect, the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use in for preventinga disorder associated with mitochondrial dysfunction.

Another aspect of the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use in for reducingthe risk of a disorder associated with mitochondrial dysfunction.

In another aspect, the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use in forameliorating a disorder associated with mitochondrial dysfunction.

Another aspect of the present disclosure relates to a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof for use in promotingoxidative metabolism.

In another aspect, the present disclosure relates to a method oftreating, preventing, ameliorating or reducing the risk of a disease ordisorder associated with α-amino-β-carboxymuconate-ε-semialdehydedecarboxylase (ACMSD) dysfunction, comprising administering to a subjectin need thereof, a therapeutically effective amount of compound havingthe following Formulae:

or a pharmaceutically acceptable salt thereof.

In a further aspect, the present disclosure relates to a method oftreating, preventing, ameliorating or reducing the risk of a disease ordisorder associated with α-amino-β-carboxymuconate-ε-semialdehydedecarboxylase (ACMSD) dysfunction, comprising administering to a subjectin need thereof, a therapeutically effective amount of compound havingthe following Formulae:

or a pharmaceutically acceptable salt thereof.

Hence, the disclosure also relates to a compound of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, as defined herein, for use asa medicament.

Another aspect of the present disclosure relates to the use of acompound having the one of the following Formula:

or a pharmaceutically acceptable salt thereof.

in the manufacture of a medicament for treating, preventing,ameliorating or reducing the risk of a disease or disorder associatedwith α-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction.

Another aspect of the present disclosure relates to the use of acompound having the one of the following Formula:

or a pharmaceutically acceptable salt thereof

in the manufacture of a medicament for treating, preventing,ameliorating or reducing the risk of a disease or disorder associatedwith α-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction.

In another aspect, the present disclosure relates to a compound havingthe one of the following Formula:

or a pharmaceutically acceptable salt thereof,

for use as a medicament for treating, preventing, ameliorating orreducing the risk of a disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction.

In another aspect, the present disclosure relates to a compound havingthe one of the following Formula:

or a pharmaceutically acceptable salt thereof,

for use as a medicament for treating, preventing, ameliorating orreducing the risk of a disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction.

Another aspect of the present disclosure relates to a compound havingthe one of the following Formula:

or a pharmaceutically acceptable salt thereof

for use in treating, preventing, ameliorating or reducing the risk of adisease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction.

In another aspect, the present disclosure relates to a compound havingthe one of the following Formula:

or a pharmaceutically acceptable salt thereof

for use in treating, preventing, ameliorating or reducing the risk of adisease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction.

As used herein, “treating” or “treat” describes the management and careof a patient for the purpose of reversing, inhibiting, or combating adisease, condition, or disorder and includes the administration of acompound of the present disclosure (i.e., a compound of Formula (I),Formula (Ia), Formula (Ib), Formula (II), or Formula (III)), or apharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, to reverse the disease, condition, or disorder,eliminate the disease, condition, or disorder, or inhibit the process ofthe disease, condition, or disorder.

A compound of the present disclosure (i.e., a compound of Formula (I),Formula (Ia), Formula (Ib), Formula (II), or Formula (III)), or apharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, can also be used to prevent a disease, condition, ordisorder or one or more symptoms of such disease, condition, ordisorder. As used herein, “preventing” or “prevent” describes reducingor eliminating the onset of the symptoms or complications of thedisease, condition, or disorder.

A compound of the present disclosure (i.e., a compound of Formula (I),Formula (Ia), Formula (Ib), Formula (II), or Formula (III)), or apharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, can also be used to alleviate one or more symptoms ofsuch disease, condition, or disorder. As used herein, the term“alleviate” is meant to describe a process by which the severity of asign or symptom of a disorder is decreased. Importantly, a sign orsymptom can be alleviated without being eliminated. Preferably treatmentis curative or ameliorating.

Methods for the Preparation of Compounds of Formulae (I), (Ia), (Ib),(II) and (III)

The compounds of the present disclosure (e.g., compounds of Formula (I),Formula (Ia), Formula (Ib), Formula (II), and Formula (II)) can beprepared in a number of ways well known to those skilled in the art oforganic synthesis. By way of example, compounds of the presentdisclosure can be synthesized using the methods described below,together with synthetic methods known in the art of synthetic organicchemistry, or variations thereon as appreciated by those skilled in theart. Preferred methods include but are not limited to those methodsdescribed below. The final products of the reactions described hereinmay be isolated by conventional techniques, e.g., by extraction,crystallisation, distillation, chromatography, etc.

Compounds of the present disclosure can be synthesized by following thesteps outlined in General Scheme A to E which comprise differentsequences of assembling intermediates Ia-Ih and Ij-Io. Startingmaterials are either commercially available or made by known proceduresin the reported literature or as illustrated. Useful steps that may beused in the preparation steps of the compounds will be known to theskilled person. The method below is given as a non-limiting example onhow the compounds may be prepared.

wherein R¹, R^(c), R^(d), and L are defined as in Formula (I).

The general way of preparing compounds of Formula (I) by usingintermediates Ia, and Ib is outlined in General Scheme A. Coupling of Iawith Ib using a base, i.e., potassium carbonate (K₂CO₃), in a solvent,i.e., acetonitrile (CH₃CN), optionally at elevated temperature providesthe desired produce of Formula (I). Bases that can be used include, butare not limited to, sodium carbonate (Na₂CO₃), potassium carbonate(K₂CO₃), N,N-diisopropylethylamine (DIPEA) and triethylamine Solventsused in the coupling reaction can be polar or non-polar solvents. Forexample, the solvent can be acetonitrile (CH₃CN), acetone, ordimethylsulfoxide (DMSO).

wherein X is a good leaving group, i.e., Cl, Br, —SCH₃, or S(O)₂CH₃, andR¹, R², R^(c), R^(d), and p are defined as in Formula (I).

Alternatively, compounds of Formula (I) can be prepared usingintermediates Ic and Id as outlined in General Scheme B. Amination ofIntermediate Ic with Ie using a base, i.e., sodium hydroxide (NaOH),potassium hydroxide (KOH), etc., in a solvent, i.e., methanol (MeOH),ethanol (EtOH), water (H₂O), etc., provides compounds of Formula (I).

wherein X is a good leaving group, i.e., Cl, Br, —SCH₃, or S(O)₂CH₃, andR¹, R², R^(c), R^(d), and p are defined as in Formula (I).

Compounds of Formula (I) can also be prepared using intermediates Ie andIf as outlined in General Scheme C. Amination of Intermediate Ie with Ifusing a base, i.e., sodium hydroxide (NaOH), potassium hydroxide (KOH),etc., in a solvent, i.e., methanol (MeOH), ethanol (EtOH), water (H₂O),etc., provides compounds of Formula (I).

wherein and R¹, R^(c), and R^(d) are defined as in Formula (I).

Alternatively, compounds of Formula (I) can also be prepared usingintermediates Ig, Ih, Ij, Ik, and Im as outlined in General Scheme D.Olefination of intermediate Ig using a base i.e., potassium carbonate(K₂CO₃) and diethyl (cyanomethyl)phosphonate in a solvent, i.e.,tetrahydrofuran (THF), water (H₂O), optionally at an elevatedtemperature provides Intermediate Ih. Hydrogenation of Ih using a metalcatalyst, i.e., palladium on carbon (Pd/C), platinum dioxide (PtO₂),etc, and hydrogen (H₂) gas in a solvent, i.e., ethanol (EtOH) and/ortetrahydrofuran (THF), provides Intermediate Ij. Intermediate Ik isobtained by treating Intermediate Ij with an acid, i.e., hydrochloricacid (HCl) in a solvent, i.e., ethanol (EtOH), dichloromethane (CH₂Cl₂),etc., and then subsequent treatment with a base, i.e., ammonia (NH₃).Cyclization of Intermediate Ik and Im using a base, i.e., sodiumhydroxide (NaOH), potassium hydroxide (KOH), etc., in a solvent, i.e.,dimethylacetamide (DMA), optionally at elevated temperature providescompounds of Formula (I).

wherein and R¹, R^(c), and R^(d) are defined as in Formula (I).

Alternatively, compounds of Formula (I) can be prepared usingintermediates In and Io as outlined in General Scheme D. Acylation ofIntermediate In with Io using a base, i.e., sodium hydroxide (NaOH),potassium hydroxide (KOH), etc., in a solvent, i.e., methanol (MeOH),ethanol (EtOH), water (H₂O), etc., provides compounds of Formula (I).

A mixture of enantiomers, diastereomers, cis/trans isomers resultingfrom the process described above can be separated into their singlecomponents by chiral salt technique, chromatography using normal phase,reverse phase or chiral column, depending on the nature of theseparation.

It should be understood that in the description and formula shown above,the various groups R¹, R², X, L, Y, R^(a), R^(b), R^(c), R^(d), R^(e),R^(f), R^(x), R^(y), R^(z), m, n, p, q, r and other variables are asdefined herein above, except where otherwise indicated. Furthermore, forsynthetic purposes, the compounds of General Schemes A-E are mererepresentative with elected radicals to illustrate the general syntheticmethodology of the compounds of Formula (I) as defined herein.

Biological Assays and Animals Studies

Method of Screening ACMSD1 Inhibition

The activity of compounds as inhibitors of ACMSD1 is determined in aspectrophotometrical in vitro assay. The pre-assay mixture is incubatedand a compound of Formula (I), Formula (Ia), Formula (Ib), Formula (II),or Formula (III), or a pharmaceutically acceptable salt thereof, andACMSD1 solution is then added. The effect of ACMS concentration on theenzyme activity is investigated by varying 3-hydroxyanthranilic acid(3OH-HA) concentration in the pre-assay mixture. Kinetic parameters arecalculated from the initial velocity data using a Lineweaver-Burk plot.

Cellular Assay Methods

The mouse hepatocytes cell lines are grown and plated. The cells aremaintained in culture at 37° C. and once the cells are attached,different concentrations of a compound of Formula (I), Formula (Ia),Formula (Ib), Formula (II), or Formula (III), or a pharmaceuticallyacceptable salt thereof, or DMSO are added. Primary hepatocytes areharvested about 24 hrs later.

Determination of ACMSD-1 Modulation in HEK293T Cells.

HEK293T cells are seeded and transfected to transiently express ACMSD.The cells are then stimulated with different concentrations of Compound1, and then lysed to measure the ACMSD activity in aspectrophotometrical in vitro assay. The amount of the whole proteincontent in cell lysates is detected by Bradford analysis and used to getthe specificity activity of the enzyme normalized in all samples.

Determination of NAD⁺ Content in Human Primary Hepatocytes

Primary hepatocytes are treated with different concentrations of acompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof, or MEHP(control) after seeding. The compound is replaced every 24 hours, andthen cells are directly harvested and lysed to detect NAD⁺ contentthrough LC MS/MS (liquid chromatography mass spectrometry/massspectroscopy).

Modulation of SOD2 Activity in AML12 Cells and Murine PrimaryHepatocytes

Primary hepatocytes or AML-12 cells are lysed and total proteinconcentration is determined using the Bradford assay. SOD2 activity isdetermined at indicated times after treatment with a compound of Formula(I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, using a SOD Assay Kit.Absorbance is determined and results are expressed in U/ml/mg of proteinaccording to the standard curve and measured protein concentration.

Determination of NAD⁺ Content in Murine Primary Hepatocytes

NAD⁺ is extracted using acidic extraction method and samples arecollected and homogenized. After insoluble protein parts are pelleted,the samples are separated by high-performance liquid chromatography(HPLC) and analyzed by mass-spectrometry. The proteins in the pellet arequantified by Bradford assay and are used for normalization.

RNA Preparation and RT-qPCR Analysis of ACMSD and SIRT1-Regulated Genesin Cells,

Cells (AML-12, Hepa-1.6, HEK-293, primary human and murine hepatocytes)are treated with different concentrations of a compound of Formula (I),Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof and the gene expression ofACMSD, Pgc1a, Sod1, and Sod2 (MnSOD) is determined using RT-qPCR. TotalRNA is extracted from cells and the extracted RNA is treated with DNaseand used for reverse transcription (RT).

Modulation of Caspase 3/7 Activity in MDCK Cells

MDCK cells are cultured in base medium to a final concentration of 10%.Cells are plated into 96 wells and 24 hours after cell plating themedium is changed with fresh medium supplemented with 1% FBS. Cisplatinis then used to induce cell injury. Different concentrations of Formula(I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof (in DMSO) are added incombination with cisplatin or prior to adding cisplatin. Caspase 3/7activity (Promega) is determined according to standard procedures usinga luminescent signal readout on a plate reader. Eachexperiment/condition is performed in triplicate. Caspase activity isanalyzed as percentage effect normalized to the cisplatin alone andvehicle treated cells.

Cytotoxicity and hERG Screening

HePG2 and AML-12 cells are seeded and a dose-response of the compound isperformed at various concentrations. Cells are stimulated and thesupernatant is used to perform LDH release as a measure of necrosiswhile the cells are lysed to detect ATP levels for determining cellviability.

The Predictor hERG assay kit is stably transfected with hERG potassiumchannel and a high-affinity red fluorescent hERG channel ligand and isused for the determination of hERG channel affinity binding of compoundsof Formula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula(III), or a pharmaceutically acceptable salt thereof. Compounds thatbind to the hERG channel protein (competitors) are identified by theirability to displace the tracer which results in a lower fluorescencepolarization.

C. elegans Experiments—ACMSD1 Silencing, Lifespan Assays, MobilityAssessment and GFP Quantification

ACMSD1 silencing: Bacterial feeding RNAi experiments to determine theeffects of downregulation or silencing of acmsd-1 on gene expression andsurvival are carried out in the nematode Caenorhabditis elegans (C.elegans). The clones used for the bacterial feeding experiments areacmsd-1, SIR-2.1 and DAF-16. Total RNA is extracted from cells and theextracted RNA is treated with DNase, and used for reverse transcription(RT).

Worms are grown on NGM agar plates additionally containing Carbenicillinand IPTG and seeded with bacterial cultures. After RNAi treatment, wormsare transferred to plates containing paraquat and seeded with RNAibacteria. Control animals are grown on RNAi bacteria containing an emptyvector (control) and then transferred to plates containing paraquat andseeded with RNAi bacteria. Quantification of gene expression of sod-3 atmRNA levels and protein levels using RT-qPCR and survival analyses areperformed. The movement of worms is recorded at days 1, 3, and 5 ofadulthood.

Anti-diabetic Effects Studies in C57BL/6J and KK-Ay Mice

Mice are fed with regular chow or a high fat diet (HFD). A compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof, is dosed daily and bloodand tissues are harvested for RNA isolation, lipid measurements andhistology. Oxygen consumption is measured and histological analysis andtransmission electron microscopy are performed. An oral glucosetolerance test and an intraperitoneal insulin tolerance test are alsoperformed to quantify glucose and to measure plasma insulinconcentrations.

Anti-diabetic and Anti-obesity Studies in db/db Mice with LepR Mutation

Animals are fed a high-fat diet (HFD). For subchronic intervention, theanimals are treated once/day with a compound of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, for 14 days. Blood samples arecollected and glucose concentrations of each blood sample aredetermined. For acute intervention, initial blood samples are collectedand then compounds of Formula (I), Formula (Ia), Formula (Ib), Formula(II), or Formula (III), or a pharmaceutically acceptable salt thereof,are administered. Diet-access is then restricted, and a second bloodsample is collected. The mice are subjected to an oral glucose tolerancetest and blood glucose concentrations are determined.

For the euglycemic-hyperinsulinemic clamps assay, the animals receive aprimed-continuous [3-³H]glucose infusion and a blood sample is thencollected to determine plasma insulin, glucose and [3-³H]glucoseconcentrations and to calculate basal endogenous glucose appearancerates. The mice then receive vehicle or a compound of Formula (I),Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, via gavage. Subsequently, theanimals receive a [3-³H]glucose infusion containing insulin causing amoderate net-increase in plasma insulin concentrations. Blood glucoseconcentrations are measured and target glycemia is established byadjusting the rate of glucose infusion. 2-deoxy-D-[1-¹⁴C] glucose isthen given intravenously and blood samples are collected. The mice arethen sacrificed. Gastrocnemius muscle and epididymal adipose tissue arecollected and plasma [³H]— and [¹⁴C]-radioactivity is determined indeproteinized plasma.

Body weights are assessed and brown adipose tissue (BAT) and gonadalwhite adipose tissue (WAT) are dissected and weighed. Volume oxygen(VO₂) and volume carbon dioxide production (VCO₂) are measured and arereported as average VO₂ per hour normalized to body weight (mL/h/kg).Activity counts by infrared beam interruptions and food intake aresimultaneously measured.

Non-alcoholic Fatty Liver Disease (NAFLD) and Non-alcoholicSteatohepatitis (NASH) Studies in Male C57BL/6J Mice

Mice are fed a ‘Western’ HF-HSD (high fat-high sucrose diet) or normalchow diet (NCD) as control. The animals are then treated with a compoundof Formula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula(III), or a pharmaceutically acceptable salt thereof, for 4, 12 or 20weeks, and then sacrificed. Body weight and food intake are monitoredweekly and total fat mass is analysed. An intraperitoneal glucosetolerance test (IPGTT) is also performed and tail vein glucose levelsare measured after glucose administration. Insulin resistance iscalculated using the Homeostasis Model of Insulin Resistance. The miceare then sacrificed by blood sampling via cardiac puncture. Plasma isobtained and tissues were collected together with the plasma for furtherbiochemical and molecular analyses or for histological analysis.

Non-alcoholic Fatty Liver Disease (NAFLD) and Non-alcoholicSteatohepatitis (NASH) Studies in Methionine and Choline Deficient Mice

Mice weighing 25 g are either fed a methionine- and choline-deficientdiet (MCD to induce NASH) or chow diet (as a control). Animalexperiments and evaluation of NAFLD and NASH are conducted as describedabove in for C57BL/6J mice fed the high fat and high sucrose diet.

Atherosclerosis Studies in High Cholesterol Fed LDL-R Knockout Mice

LDL-R knockout (KO) mice are sacrificed about 12 weeks after theinitiation of the atherogenic diet, after which the heart and aorta areperfused with PBS and subsequently fixed. Atherosclerosis andbiochemistry parameters are measured with the appropriate commerciallyavailable kits. For the in vivo lipopolysaccharide (LPS) study, mice areintraperitoneally injected with LPS, and blood is taken from the tailvein. TNFα levels are quantified with a Mouse TNFα ELISA assay. Bloodcell counts are determined.

Inherited Mitochondrial Disease Studies in Sco2^(KO/KI) Mice

Compounds of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III) are dissolved in water and added to a standard powder dietat the appropriate concentration. The diet supply is changed every threedays and administered ad libitum for one month. Tissues are collectedfor histological analysis. For the muscle quadriceps samples, thespectrophotometric activity of cI, cII, cIII, and cIV, as well as CS, ismeasured. NAD⁺ is extracted from tissues using acidic and alkalineextraction methods, respectively, and analysed with mass spectrometry.

Inherited Mitochondrial Disease Studies in Deletor Mice

Deletor and WT male mice are administered either chow diet (CD) or acompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III) admixed with the CD. The mice are regularly monitored forweight, food consumption, and physical endurance and their exercisecapability is measured. Oxygen consumption and carbon dioxideproduction, as well as spontaneous moving and feeding activities, arerecorded. Tissue sections are collected and prepared from thequadriceps, liver, and BAT. Frozen sections from quadriceps are assayedfor in situ histochemical COX and succinate dehydrogenase (SDH)activities, crista content in both BAT and muscle is determined fromelectron micrographs and skeletal muscle samples are analysed forcitrate synthase activity.

Kidney Disease Studies

C57BL/6J WT mice are fed a standard commercial diet and divided intofour groups: control; cisplatin; a compound of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, and cisplatin; and a compoundof Formula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula(III), or a pharmaceutically acceptable salt thereof, alone. The miceare sacrificed and tissue samples and serum are collected. Serumcreatinine and BUN levels are measured and the proinflammatory cytokinesTNF-α, IL-1b, and IL-6 from serum or homogenates from kidney tissue arequantified. Mouse kidneys are collected and stained for analysis.Tubular damage is examined and scored based on the percentage ofcortical tubular necrosis. Neutrophil infiltration is quantitativelyassessed on stained tissue by counting the number of neutrophils perhigh-power field.

Alternatively, C57BL/6J WT mice are numbered and kept in acclimatizationfor a period and then randomized into different treatment groups basedon their body weight. Different groups are maintained on a specifieddiet for a period of time. Body weight measurements are taken and foodconsumption is evaluated. Blood is collected by retro-orbital punctureunder mild anesthesia and used for analysis of basal blood urea nitrogenlevels (BUN).

Mice are anesthetized and placed on a surgical platform. Both kidneysare exposed through incisions and renal pedicles are occluded usingvascular clamps. The clamp is then removed and the surgical site issutured. The sham-operated group is subjected to similar surgicalprocedures, except that the occluding clamp is not applied. Animals aremonitored until recovery from anesthesia and returned to their homecage. Animals are observed every day for general clinical signs andsymptoms and mortality.

One day prior to termination, animals are individually housed inmetabolic cages and urine is collected for estimation of urea,creatinine, sodium and potassium. Blood is also collected by retroorbital puncture under mild anesthesia and plasma is used for analysisof blood urea nitrogen levels (BUN) and serum creatinine. Animals arethen euthanized and organs are collected. One kidney is fixed and theother is flash frozen and used for the estimation of lipid peroxidation,GSH, MPO and SOD levels.

Ischemia/Reperfusion-induced Acute Kidney Injury Studies

CD-1 (ICR) mice are treated with a compound of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, by oral gavage once per day.CD-1 mice are divided into four groups: (1) young mice with sham injury;(2) young mice with ischemic/reperfusion (I/R) injury; (3) adult micewith sham and (4) adult mice with I/R injury. An additional 27 adultmice are randomized into two groups: mice receiving a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof, and mice receiving thevehicle as a control. The serum creatinine level is measured and BUNmeasurements are recorded. Renal tissue is then evaluated and tubularinjury is scored.

Determination of the Effects on FoxO1 Phosphorylation Levels

AML-12 cells are treated with different concentrations of a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof. Cells are then lysed, andanalyzed by SDS-PAGE/western blot. Blocking and antibody incubations arethen done and each protein present is detected with its specificantibody.

Inhibitory Effect

The present disclosure also relates to a compound of Formula (I),Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, as defined herein, in a methodfor inhibiting the activity of ACMSD. The method includes contacting acell with a compound of Formula (I), Formula (Ia), Formula (Ib), Formula(II), or Formula (III), or a pharmaceutically acceptable salt thereof.In a related embodiment, the method further provides that the compoundis present in an amount effective to produce a concentration sufficientto selectively inhibit ACMSD in the cell.

Thus, preferably in an assay for ACMSD inhibition (i.e., an ACMSD assaydescribed herein, e.g., Example 29, or an ACMSD assays known in theliterature), the preferred compounds of Formula (I), Formula (Ia),Formula (Ib), Formula (II), or Formula (III), or a pharmaceuticallyacceptable salt thereof, are compounds capable of reducing or preferablyinhibiting ACMSD and increasing NAD⁺ levels and/or activating SIRTs andthe downstream targets of SIRTs, such as PGC-1α, FoxO1 and/or SOD.Preferably, said inhibition is determined as the IC₅₀ of said compoundof Formula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula(III) with respect to said ACMSD inhibition assay. Preferred compoundsof Formula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula(III), or a pharmaceutically acceptable salt thereof, have an IC₅₀ at orbelow 1 μM, more preferably less than 300 nM, for example less than 100nM, such as less than 50 nM with respect to inhibition of ACMSD.

Pharmaceutically Acceptable Salts

The compound of Formula (I), Formula (Ia), Formula (Ib), Formula (II),or Formula (III) may be provided in any form suitable for the intendedadministration, in particular including pharmaceutically acceptablesalts, solvates and prodrugs of the compound of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III).

Pharmaceutically acceptable salts refer to salts of the compounds ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III)which are considered to be acceptable for clinical and/or veterinaryuse. Typical pharmaceutically acceptable salts include those saltsprepared by reaction of the compounds of Formula (I), Formula (Ia),Formula (Ib), Formula (II), or Formula (III) and a mineral or organicacid or an organic or inorganic base. Such salts are known as acidaddition salts and base addition salts, respectively. It will berecognized that the particular counter-ion forming a part of any salt isnot of a critical nature, so long as the salt as a whole ispharmaceutically acceptable and as long as the counter-ion does notcontribute undesired qualities to the salt as a whole. These salts maybe prepared by methods known to the skilled person. Pharmaceuticallyacceptable salts are, e.g., those described and discussed in Remington'sPharmaceutical Sciences, 17. Ed. Alfonso R. Gennaro (Ed.), MackPublishing Company, Easton, Pa., U.S.A., 1985 and more recent editionsand in Encyclopedia of Pharmaceutical Technology.

Examples of pharmaceutically acceptable addition salts include acidaddition salts formed with inorganic acids, e.g., hydrochloric,hydrobromic, sulfuric, nitric, hydroiodic, metaphosphoric, or phosphoricacid; and organic acids e.g., succinic, maleic, acetic, fumaric, citric,tartaric, benzoic, trifluoroacetic, malic, lactic, formic, propionic,glycolic, gluconic, camphorsulfuric, isothionic, mucic, gentisic,isonicotinic, saccharic, glucuronic, furoic, glutamic, ascorbic,anthranilic, salicylic, phenylacetic, mandelic, embonic (pamoic),ethanesulfonic, pantothenic, stearic, sulfinilic, alginic andgalacturonic acid; and arylsulfonic, for example benzenesulfonic,p-toluenesulfonic, methanesulfonic or naphthalenesulfonic acid; and baseaddition salts formed with alkali metals and alkaline earth metals andorganic bases such as N,N-dibenzylethylenediamine, chloroprocaine,choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine),lysine and procaine; and internally formed salts. It should beunderstood that all references to pharmaceutically acceptable saltsinclude solvent addition forms (solvates) or crystal forms (polymorphs)as defined herein, of the same salt.

The compound of Formula (I), Formula (Ia), Formula (Ib), Formula (II),or Formula (III), or a pharmaceutically acceptable salt thereof, may beprovided in dissoluble or indissoluble forms together with apharmaceutically acceptable solvent such as water, ethanol, and thelike. Dissoluble forms may also include hydrated forms such as themono-hydrate, the dihydrate, the hemihydrate, the trihydrate, thetetrahydrate, and the like.

The compound of Formula (I), Formula (Ia), Formula (Ib), Formula (II),or Formula (III), or a pharmaceutically acceptable salt thereof, may beprovided as a prodrug. The term “prodrug” used herein is intended tomean a compound which—upon exposure to certain physiologicalconditions—will liberate the compound of Formula (I), Formula (Ia),Formula (Ib), Formula (II), or Formula (III), or a pharmaceuticallyacceptable salt thereof, which then will be able to exhibit the desiredbiological action. A typical example is a labile carbamate of an amine.

Since prodrugs are known to enhance numerous desirable qualities ofpharmaceuticals (e.g., solubility, bioavailability, manufacturing,etc.), the compounds of the present disclosure can be delivered inprodrug form. Thus, the present disclosure is intended to cover prodrugsof the presently claimed compounds, methods of delivering the same andcompositions containing the same. “Prodrugs” are intended to include anycovalently bonded carriers that release an active parent drug of thepresent disclosure in vivo when such prodrug is administered to asubject. Prodrugs in the present disclosure are prepared by modifyingfunctional groups present in the compound in such a way that themodifications are cleaved, either in routine manipulation or in vivo, tothe parent compound. Prodrugs include compounds of the presentdisclosure wherein a hydroxy, amino, sulfhydryl, carboxy or carbonylgroup is bonded to any group that may be cleaved in vivo to form a freehydroxyl, free amino, free sulfhydryl, free carboxy or free carbonylgroup, respectively.

Examples of prodrugs include, but are not limited to, esters (e.g.,acetate, dialkylaminoacetates, formates, phosphates, sulfates andbenzoate derivatives) and carbamates (e.g., N,N-dimethylaminocarbonyl)of hydroxy functional groups, esters (e.g., C₁₋₆ alkyl esters, e.g.,methyl esters, ethyl esters, 2-propyl esters, phenyl esters,2-aminoethyl esters, morpholinoethanol esters, etc.) of carboxylfunctional groups, N-acyl derivatives (e.g., N-acetyl) N-Mannich bases,Schiff bases and enaminones of amino functional groups, oximes, acetals,ketals and enol esters of ketone and aldehyde functional groups incompounds of the disclosure, and the like. See Bundegaard, H., Design ofProdrugs, p1-92, Elesevier, N.Y.-Oxford (1985).

The compounds, or pharmaceutically acceptable salts, esters or prodrugsthereof, are administered orally, nasally, transdermally, pulmonary,inhalationally, buccally, sublingually, intraperintoneally,subcutaneously, intramuscularly, intravenously, rectally,intrapleurally, intrathecally and parenterally. In one embodiment, thecompound is administered orally. One skilled in the art will recognizethe advantages of certain routes of administration.

The dosage regimen utilizing the compounds is selected in accordancewith a variety of factors including type, species, age, weight, sex andmedical condition of the patient; the severity of the condition to betreated; the route of administration; the renal and hepatic function ofthe patient; and the particular compound or salt thereof employed. Anordinarily skilled physician or veterinarian can readily determine andprescribe the effective amount of the drug required to prevent, counteror arrest the progress of the condition.

Techniques for formulation and administration of the disclosed compoundsof the disclosure can be found in Remington: the Science and Practice ofPharmacy, 19^(th) edition, Mack Publishing Co., Easton, Pa. (1995). Inan embodiment, the compounds described herein, and the pharmaceuticallyacceptable salts thereof, are used in pharmaceutical preparations incombination with a pharmaceutically acceptable carrier or diluent.Suitable pharmaceutically acceptable carriers include inert solidfillers or diluents and sterile aqueous or organic solutions. Thecompounds will be present in such pharmaceutical compositions in amountssufficient to provide the desired dosage amount in the range describedherein.

In one aspect of this disclosure, there is provided a pharmaceuticalcomposition comprising at, as an active ingredient, at least onecompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof, as definedherein, and optionally one or more pharmaceutically acceptableexcipients, diluents and/or carriers. The compounds of Formula (I),Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, may be administered alone orin combination with pharmaceutically acceptable carriers, diluents orexcipients, in either single or multiple doses. Suitablepharmaceutically acceptable carriers, diluents and excipients includeinert solid diluents or fillers, sterile aqueous solutions and variousorganic solvents.

A “pharmaceutical composition” is a formulation containing the compoundsof the present disclosure in a form suitable for administration to asubject. The pharmaceutical compositions may be formulated withpharmaceutically acceptable carriers or diluents as well as any otherknown adjuvants and excipients in accordance with conventionaltechniques such as those disclosed in Remington: The Science andPractice of Pharmacy, 21st Edition, 2000, Lippincott Williams & Wilkins.

As used herein, the phrase “pharmaceutically acceptable” refers to thosecompounds, materials, compositions, carriers, and/or dosage forms whichare, within the scope of sound medical judgment, suitable for use incontact with the tissues of human beings and animals without excessivetoxicity, irritation, allergic response, or other problem orcomplication, commensurate with a reasonable benefit/risk ratio.

“Pharmaceutically acceptable excipient” means an excipient that isuseful in preparing a pharmaceutical composition that is generally safe,non-toxic and neither biologically nor otherwise undesirable, andincludes excipient that is acceptable for veterinary use as well ashuman pharmaceutical use. A “pharmaceutically acceptable excipient” asused in the specification and claims includes both one and more than onesuch excipient.

The pharmaceutical compositions formed by combining a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof, as defined herein, withpharmaceutically acceptable carriers, diluents or excipients can bereadily administered in a variety of dosage forms such as tablets,powders, lozenges, syrups, suppositories, injectable solutions and thelike. In powders, the carrier is a finely divided solid such as talc orstarch which is in a mixture with the finely divided active component.In tablets, the active component is mixed with the carrier having thenecessary binding properties in suitable proportions and compacted inthe shape and size desired.

The pharmaceutical compositions may be specifically prepared foradministration by any suitable route such as the oral and parenteral(including subcutaneous, intramuscular, intrathecal, intravenous andintradermal) route. It will be appreciated that the preferred route willdepend on the general condition and age of the subject to be treated,the nature of the condition to be treated and the active ingredientchosen.

Pharmaceutical compositions for oral administration include solid dosageforms such as capsules, tablets, dragees, pills, lozenges, powders, andgranules. Where appropriate, they can be prepared with coatings such asenteric coatings or they can be prepared so as to provide controlledrelease of the active ingredient such as sustained or prolonged releaseaccording to methods well known in the art.

For oral administration in the form of a tablet or capsule, a compoundof Formula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula(III), or a pharmaceutically acceptable salt thereof, as defined herein,may suitably be combined with an oral, non-toxic, pharmaceuticallyacceptable carrier such as ethanol, glycerol, water, or the like.Furthermore, suitable binders, lubricants, disintegrating agents,flavouring agents, and colourants may be added to the mixture, asappropriate. Suitable binders include, e.g., lactose, glucose, starch,gelatin, acacia gum, tragacanth gum, sodium alginate,carboxymethylcellulose, polyethylene glycol, waxes, or the like.Lubricants include, e.g., sodium oleate, sodium stearate, magnesiumstearate, sodium benzoate, sodium acetate, sodium chloride, or the like.Disintegrating agents include, e.g., starch, methyl cellulose, agar,bentonite, xanthan gum, sodium starch glycolate, crospovidone,croscarmellose sodium, or the like. Additional excipients for capsulesinclude macrogels or lipids.

For the preparation of solid compositions such as tablets, the activecompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof, is mixedwith one or more excipients, such as the ones described above, and otherpharmaceutical diluents such as water to make a solid pre-formulationcomposition containing a homogenous mixture of a compound of Formula(I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof. The term “homogenous” isunderstood to mean that the compound of Formula (I), Formula (Ia),Formula (Ib), Formula (II), or Formula (III), or a pharmaceuticallyacceptable salt thereof, is dispersed evenly throughout the compositionso that the composition may readily be subdivided into equally effectiveunit dosage forms such as tablets or capsules.

Liquid compositions for either oral or parenteral administration of thecompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof, include,e.g., aqueous solutions, syrups, elixirs, aqueous or oil suspensions andemulsion with edible oils such as cottonseed oil, sesame oil, coconutoil, or peanut oil. Suitable dispersing or suspending agents for aqueoussuspensions include synthetic or natural gums such as tragacanth,alginate, acacia, dextran, sodium carboxymethylcellulose, gelatin,methylcellulose, or polyvinylpyrrolidone.

Pharmaceutical compositions for parenteral administration includesterile aqueous and non-aqueous injectable solutions, dispersions,suspensions or emulsions as well as sterile powders to be reconstitutedin sterile injectable solutions or dispersions prior to use.

For intravenous administration, suitable carriers include physiologicalsaline, bacteriostatic water, Cremophor EL™ (BASF, Parsippany, N.J.) orphosphate buffered saline (PBS). In all cases, the composition must besterile and should be fluid to the extent that easy syringeabilityexists. It must be stable under the conditions of manufacture andstorage and must be preserved against the contaminating action ofmicroorganisms such as bacteria and fungi. The carrier can be a solventor dispersion medium containing, for example, water, ethanol, polyol(for example, glycerol, propylene glycol, and liquid polyethyleneglycol, and the like), and suitable mixtures thereof. The properfluidity can be maintained, for example, by the use of a coating such aslecithin, by the maintenance of the required particle size in the caseof dispersion and by the use of surfactants. Prevention of the action ofmicroorganisms can be achieved by various antibacterial and antifungalagents, for example, parabens, chlorobutanol, phenol, ascorbic acid,thimerosal, and the like. In many cases, it will be preferable toinclude isotonic agents, for example, sugars, polyalcohols such asmanitol, sorbitol, and sodium chloride in the composition. Prolongedabsorption of the injectable compositions can be brought about byincluding in the composition an agent which delays absorption, forexample, aluminum monostearate and gelatin.

The preparation of all these solutions under sterile conditions isreadily accomplished by standard pharmaceutical techniques well known tothose skilled in the art.

For example, sterile injectable solutions can be prepared byincorporating the active compound in the required amount in anappropriate solvent with one or a combination of ingredients enumeratedabove, as required, followed by filtered sterilization. Generally,dispersions are prepared by incorporating the active compound into asterile vehicle that contains a basic dispersion medium and the requiredother ingredients from those enumerated above. In the case of sterilepowders for the preparation of sterile injectable solutions, methods ofpreparation are vacuum drying and freeze-drying that yields a powder ofthe active ingredient plus any additional desired ingredient from apreviously sterile-filtered solution thereof. Depot injectablecompositions are also contemplated as being within the scope of thepresent disclosure.

For parenteral administration, solutions containing a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof, in sesame or peanut oil,aqueous propylene glycol, or in sterile aqueous solution may beemployed. Such aqueous solutions should be suitably buffered ifnecessary and the liquid diluent first rendered isotonic with sufficientsaline or glucose. These particular aqueous solutions are especiallysuitable for intravenous, intramuscular, subcutaneous andintraperitoneal administration. The oily solutions are suitable forintra-articular, intramuscular and subcutaneous injection purposes.

In addition to the aforementioned ingredients, the compositions of acompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof, mayinclude one or more additional ingredients such as diluents, buffers,flavouring agents, colourant, surface active agents, thickeners,preservatives, e.g., methyl hydroxybenzoate (including anti-oxidants),emulsifying agents and the like.

The term “therapeutically effective amount”, as used herein, refers toan amount of a pharmaceutical agent to treat, ameliorate, or prevent anidentified disease, disorder, or condition, or to exhibit a detectabletherapeutic or inhibitory effect. The effect can be detected by anyassay method known in the art. The precise effective amount for asubject will depend upon the subject's body weight, size, and health;the nature and extent of the condition; and the therapeutic orcombination of therapeutics selected for administration. Therapeuticallyeffective amounts for a given situation can be determined by routineexperimentation that is within the skill and judgment of the clinician.In a preferred aspect, the disease or disorder to be treated is adisease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction.

For any compound, the therapeutically effective amount can be estimatedinitially either in cell culture assays, e.g., in cells, or in animalmodels, usually rats, mice, rabbits, dogs, or pigs. The animal model mayalso be used to determine the appropriate concentration range and routeof administration. Such information can then be used to determine usefuldoses and routes for administration in humans. Therapeutic/prophylacticefficacy and toxicity may be determined by standard pharmaceuticalprocedures in cell cultures or experimental animals, e.g., ED₅₀ (thedose therapeutically effective in 50% of the population) and LD₅₀ (thedose lethal to 50% of the population). The dose ratio between toxic andtherapeutic effects is the therapeutic index, and it can be expressed asthe ratio, LD₅₀/ED₅₀. Pharmaceutical compositions that exhibit largetherapeutic indices are preferred. The dosage may vary within this rangedepending upon the dosage form employed, sensitivity of the patient, andthe route of administration.

Dosage and administration are adjusted to provide sufficient levels ofthe active agent(s) or to maintain the desired effect. Factors which maybe taken into account include the severity of the disease state, generalhealth of the subject, age, weight, and gender of the subject, diet,time and frequency of administration, drug combination(s), reactionsensitivities, and tolerance/response to therapy. Long-actingpharmaceutical compositions may be administered every 3 to 4 days, everyweek, or once every two weeks depending on half-life and clearance rateof the particular formulation.

A suitable dosage of the compound of Formula (I), Formula (Ia), Formula(Ib), Formula (II), or Formula (III), or a pharmaceutically acceptablesalt thereof, will depend on the age and condition of the patient, theseverity of the disease to be treated and other factors well known tothe practicing physician. The compound may be administered for exampleeither orally, parenterally or topically according to different dosingschedules, e.g., daily or with intervals, such as weekly intervals. Ingeneral a single dose will be in the range from 0.01 to 500 mg/kg bodyweight, preferably from about 0.05 to 100 mg/kg body weight, morepreferably between 0.1 to 50 mg/kg body weight, and most preferablybetween 0.1 to 25 mg/kg body weight. The compound may be administered asa bolus (i.e., the entire daily dose is administered at once) or individed doses two or more times a day. Variations based on theaforementioned dosage ranges may be made by a physician of ordinaryskill taking into account known considerations such as weight, age, andcondition of the person being treated, the severity of the affliction,and the particular route of administration.

As used herein, a “subject” or “subject in need thereof” is a subjecthaving a disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction. A “subject” includes a mammal. The mammal can be e.g., anymammal, e.g., a human, primate, bird, mouse, rat, fowl, dog, cat, cow,horse, goat, camel, sheep or a pig. Preferably, the mammal is a human.

The compounds of Formula (I), Formula (Ia), Formula (Ib), Formula (II),or Formula (III), or a pharmaceutically acceptable salt thereof, mayalso be prepared in a pharmaceutical composition comprising one or morefurther active substances alone, or in combination with pharmaceuticallyacceptable carriers, diluents, or excipients in either single ormultiple doses. The suitable pharmaceutically acceptable carriers,diluents and excipients are as described herein above, and the one ormore further active substances may be any active substances, orpreferably an active substance as described in the section “combinationtreatment” herein below.

Clinical Conditions and Other Uses of Compounds

The compounds according to Formula (I), Formula (Ia), Formula (Ib),Formula (II), or Formula (III), or a pharmaceutically acceptable formthereof, compositions, medicaments, and compounds for use, as definedherein, are useful for treatment of a disease or disorder in whichα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)modulation plays a role. The compounds may be used either in human or inveterinary medicine and the patient may be any mammal, but especially ahuman. The treatment may include administering to any mammal, butespecially a human, suffering from a disease or disorder in whichα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)modulation plays a role, a therapeutically effective amount of acompound according to Formula (I), Formula (Ia), Formula (Ib), Formula(II), or Formula (III), or a pharmaceutically acceptable salt thereof,as defined herein.

The present disclosure also relates to a compound of Formula (I),Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, as defined herein, for use ina disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction, such as obesity, type II diabetes and its complications(e.g., diabetic retinopathy and nephropathy), non-alcoholic fatty liverdisease (NAFLD), non-alcoholic steatohepatitis (NASH), or chronic kidneydisease.

By the term “disease or disorder associated withα-amino-β-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD)dysfunction” is meant any disease characterized by reduced nicotinamideadenine dinucleotide (NAD⁺) expression and/or activity in at least insome instances of the disease, or a disease which is ameliorated byelevation of the levels of NAD⁺.

The methods, medicaments and compounds for use of the present disclosureare useful to treat, alleviate the symptoms of, or delay the onset of adisorder associated with aberrant mitochondrial function. Disordersassociated with aberrant mitochondrial function include, for example,metabolic disorders, neurodegenerative disorders, aging relateddisorders, and chronic inflammatory disorders. Mitochondrial disordersalso include diseases with inherited and/or acquired mitochondrialdysfunction (i.e., Charcot-Marie-tooth disease, Type 2A2, MitochondrialEncephalopathy Lactic Acidosis and Stroke (MELAS), Leigh syndrome, Barthsyndrome, and Leber's optic neuropathy), fatty acid oxidation disorders,inherited forms of deafness and blindness, and metabolic abnormalitiesinduced by exposure to toxic chemicals and/or drugs (e.g., cisplatininduced deafness).

Metabolic disorders include, for example, type II diabetes, obesity,hyperglycemia, glucose intolerance, insulin resistance (i.e.,hyperinsulinemia, metabolic syndrome, syndrome X), hypercholesterolemia,hypertension, hyperlipoproteinemia, hyperlipidemia (e.g., dyslipidemia),hypertriglylceridemia, cardiovascular disease, atherosclerosis,peripheral vascular disease, kidney disease, ketoacidosis, thromboticdisorders, nephropathy, diabetic neuropathy, diabetic retinopathy,sexual dysfunction, dermatopathy, dyspepsia, hypoglycemia, cancer, andedema.

Neurodegenerative disorders include diseases such as photoreceptordegeneration (i.e., retinitis pigmentosa), Dementia, Alzheimer'sdisease, Parkinson's disease, and Huntington's disease.

Chronic inflammatory diseases include diseases such as celiac disease,vasculitis, lupus, chronic obstructive pulmonary disease (COPD),irritable bowel disease, atherosclerosis, arthritis, and psoriasis.

Aging related disorders include diseases such as cancer, dementia,cardiovascular disease (i.e., arteriosclerosis), hypertension, diabetesmellitus (type I or type II), arthritis, cataracts, Alzheimer's disease,macular degeneration, and osteoporosis.

The subject can be suffering from or susceptible to developing ametabolic disorder. Subjects suffering from or at risk of developing ametabolic disorder are identified by methods known in the art. Forexample, diabetes can be diagnosed by measuring fasting blood glucoselevels or insulin or by glucose tolerance test. Normal adult glucoselevels are between about 60-126 mg/dl. Normal insulin levels are about 7mU/mL±3 mU. Hypertension can be diagnosed by a blood pressure readingconsistently at or above about 140/90. Cardiovascular disease can bediagnosed by measuring cholesterol levels. For example, LDL cholesterolabove about 137 or total cholesterol above about 200 is indicative ofcardiovascular disease. Hyperglycemia can be diagnosed by a bloodglucose level higher than about 10 mmol/1(180 mg/dl). Glucoseintolerance can be diagnosed by glucose levels of 140 to 199 mg per dL(7.8 to 11.0 mmol) after conducting a 75 g oral two-hour glucosetolerance test. Insulin resistance can be diagnosed by a fasting seruminsulin level of greater than approximately 60 pmol/L. Hypoglycemia canbe diagnosed by a blood glucose level lower than about 2.8 to 3.0 mmol/L(50 to 54 mg/dl). Obesity can be diagnosed, for example, by body massindex. Body mass index (BMI) is measured in kg/m² (or lb/in²×704.5).Alternatively, waist circumference (estimates fat distribution),waist-to-hip ratio (estimates fat distribution), skinfold thickness (ifmeasured at several sites, estimates fat distribution), or bioimpedance(based on principle that lean mass conducts current better than fat mass(i.e., fat mass impedes current), estimates % fat) can be measured. Theparameters for normal, overweight, or obese individuals are as follows:Underweight: BMI<18.5; Normal: BMI about 18.5 to about 24.9; Overweight:BMI=about 25 to about 29.9. Overweight individuals are characterized ashaving a waist circumference of >94 cm for men or >80 cm for women andwaist to hip ratios of ≧0.95 in men and ≧0.80 in women. Obeseindividuals are characterized as having a BMI of 30 to 34.9, beinggreater than 20% above “normal” weight for height, having a body fatpercentage >30% for women and 25% for men, and having a waistcircumference >102 cm (40 inches) for men or 88 cm (35 inches) forwomen. Individuals with severe or morbid obesity are characterized ashaving a BMI of ≧35.

The methods described herein may lead to a reduction in the severity orthe alleviation of one or more symptoms of a metabolic disorder. Forexample, symptoms of diabetes include elevated fasting blood glucoselevels, blood pressure at or above 140/90 mm/Hg; abnormal blood fatlevels, such as high-density lipoproteins (HDL) less than or equal to 35mg/dL, or triglycerides greater than or equal to 250 mg/dL(mg/dL=milligrams of glucose per deciliter of blood). Efficacy oftreatment is determined in association with any known method fordiagnosing the metabolic disorder. Alleviation of one or more symptomsof the metabolic disorder indicates that the compound confers a clinicalbenefit.

The methods of the present disclosure are useful to treat, alleviate thesymptoms of, or delay the onset of a kidney disorder. Kidney disordersinclude acute kidney injury (AKI) and chronic kidney disease (CKD).

The subject can be suffering from or susceptible to developing acutekidney injury (AKI). The acute kidney injury can be characterized by oneor more clinical criteria or conditions (i.e., an abrupt decrease in theability of the kidneys to excrete nitrogenous waste products from theblood, resulting in azotemia). Subjects suffering from or at risk ofdeveloping acute kidney injury (AKI) are identified by methods known inthe art. For example, the acute kidney injury can be characterized by anincrease in serum creatinine by at least 50% over baseline, an absoluteincrease in serum creatinine of at least 0.3 mg/dL over baseline, areduction in glomerular filtration rate of at least 25% compared tobaseline, a decrease in urine output to 0.5 ml per kilogram of bodyweight or less per hour persisting for at least 6 hours, or anycombination thereof. An acute kidney injury may be caused by ischemia,drugs or toxic agents radiocontrast media, a non-steroidalanti-inflammatory drug (NSAID), alcohol, or a chemotherapy agent),viruses, and obstruction.

The subject can be suffering from or susceptible to developing chronickidney disease (CKD). Chronic kidney disease (CKD) is defined as either(1) having kidney damage as defined by structural or functionalabnormalities of the kidney for 3 months or longer with or without adecreased glomerular filtration rate (GFR) or (2) having a GFR of lessthan 60 mL/min/1.73 m² for 3 months or longer with or without kidneydamage. Subjects suffering from or at risk of developing a chronickidney disease (CKD) are identified by methods known in the art.Structural or functional abnormalities are manifested by symptoms suchas either pathologic abnormalities or markers of kidney damage,including abnormalities identified in imaging studies or the compositionof blood or urine.

For example, CKD can be diagnosed by testing for specific marker. Forexample, markers of kidney damage include a plasma creatinineconcentration of above about 1.6 mg/dL and a blood urea nitrogen (BUN)concentration of above about 20 mg/dL. Typically, both of these markersare elevated in individuals with CKD. Additional markers of kidneydamage can include hematuria (i.e., any detectable amount of blood inthe urine), proteinuria (i.e., protein concentrations in urine aboveabout 100 mg/dL), albuminuria (i.e., albumin concentrations in urineabove about 100 m/dL), an intact parathyroid hormone (PTH) concentrationin the blood above about 150 pg/mL, or blood phosphate levels of aboveabout 4.5 mg/dL. One specific marker of kidney disease is a GFR rateabove normal (i.e., a GFR above about 90 mL/min/1.73 m²), however abelow normal GFR also indicates CKD.

The methods of the present disclosure are useful to treat, alleviate thesymptoms of, or delay the onset of non-alcoholic fatty liver disease(NAFLD) and/or non-alcoholic steatohepatitis (NASH). The subject can besuffering from or susceptible to developing non-alcoholic fatty liverdisease (NAFLD) and/or non-alcoholic steatohepatitis (NASH). Subjectssuffering from or at risk of developing a non-alcoholic fatty liverdisease (NAFLD) and/or non-alcoholic steatohepatitis (NASH) areidentified by methods known in the art. For example, NAFLD and/or NASHcan be diagnosed by liver biopsy.

Non-alcoholic fatty liver disease (NAFLD), as defined herein, is adisease with it deposition in the liver, which occurs in patients whosealcohol ingestion history is not long enough to cause liver injury.Non-alcoholic fatty liver disease (NAFLD) can be further classified intosimple fatty liver, steatohepatitis and cirrhosis. Nonalcoholicsteatohepatitis (NASH) refers to a pathology associated withinflammation, liver cell necrosis, ballooning and fibrosis. The onset ofnonalcoholic simple fatty liver is induced by fat deposition in livercells, and this fat accumulation is defined by the balance betweenincreasing factors (influx and synthesis of fats in liver cells) anddecreasing factors (catabolism of its and their release from livercells). Once damage of liver cells occurs, in addition to this fatdeposition, nonalcoholic simple fatty liver will progress tononalcoholic steatohepatitis. Nonalcoholic steatohepatitis isprogressive and may finally progress to cirrhosis and hepatocellularcarcinoma.

Combination Treatment

A compound, compositions, medicaments and compounds for use of Formula(I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, may also be used to advantagein combination with one or more other therapeutic agents. Suchtherapeutic agents include, but are not limited to other ACMSDinhibitors; anti-diabetic agents such as PPARy agonists, PPARα/γ dualagonists, PPARδ agonists, biguanides, protein tyrosine phosphatase-1B(PTP-1B), dipeptidyl peptidase IV (DPP-IV) inhibitors, sulfonylureas,meglitinides, alpha glucoside hydrolase inhibitors, alpha-amylaseinhibitors, insulin secreatagogues, A2 antagonists, insulin or insulinmimetics, glycogen phosphorylase inhibitors, GLP-1 agonists,non-thiazolidinediones, glycokinase, and 11β HSD-1 inhibitor;anti-obesity agents such as uncoupling Protein (UCP-1, UCP-2, and UCP-3)activators, β3 adrenergic receptor (β3), thyroid hormone β agonists,fatty acid synthase (PAS) inhibitors, phosphodieterase (PDE) inhibitors,lipase inhibitors, serotonin reuptake inhibitors, monoamine reuptakeinhibitors, Mc4r agonists, 5HT2c agonists, growth hormone secretagogue(GHS) agonists, CNTF derivatives, ciliary neurotrophic factors (CNTh),cholecystokinin-A (CCK-A) agonists, opioid antagonists, orexinantagonists, acyl-estrogens, leptin, NPY 5 antagonists, neuropeptide Y5(NPY5) antagonists, neuropeptide Y2 (NPY2) agonists,melanin-concentrating hormone receptor (MCHLR) antagonists andmelanin-concentrating hormone 2 receptor (MCH2R), MCH1R antagonists,neuropeptide Y1, ghrelin antagonists, cannabinoid receptor 1 (CB-1),serotonin (5HT) transport inhibitors, CCK-A agonists and histamine 3(H3) antagonist/inverse agonists; cholesterol lower agents such as3-hydroxy-3-methylglutaryl-coenzyme A (HMG CoA) reductase inhibitors,HMG-CoA synthase inhibitors, squalene epoxidase inhibitors, fibricacids, bile acid-binding resins probucol and niacin (nicotinic acid);compounds that boost NAD⁺ levels such as NAD⁺ precursors (i.e.,nicotinamide ribose (NA), nicotinamide mononucleotide (NMN), nicotinicacid (NA) and nicotinamide); and compounds that inhibit NAD⁺ consumptionsuch as PARP inhibitors and CD38 inhibitors.

PPARy agonists useful in the present disclosure include, but are notlimited to, glitazones (e.g., balaglitazone, ciglitazone, darglitazone,englitazone, isaglitazone (MCC-555), pioglitazone, rosiglitazone,troglitazone, CLX-0921, 5-BTZD, and the like); GW-0207, LG-100641,LY-300512, LY-519818, R483 (Roche), T131 (Tularik), and compoundsdisclosed in WO97/27857, 97/28115, 97/28137 and 97/27847; andpharmaceutically acceptable salts or esters thereof. PPARα/γ dualagonists useful in the present disclosure, include, but are not limitedto, CLX-0940, GW-1536, GW1929, GW-2433, KRP-297, L-796449, LR-90,MK-0767, SB 219994, and muraglitazar, and pharmaceutically acceptablesalts or esters thereof. KRP-297 is5-[(2,4-Dioxo-5-thiazolidinyl)methyl]-2-methoxy-N-[[4-(trifluoromethyl)phenyl]methyl]benzamide,and pharmaceutically acceptable salts or esters thereof. PPARδ agonistsuseful in the present disclosure include, but are not limited to, GW501516, GW 590735, and compounds disclosed in JP 10237049, and WO02/14291; and pharmaceutically acceptable salts or esters thereof.

Biguanides useful in the present disclosure include, but are not limitedto, buformin, metformin, and phenformin, and pharmaceutically acceptablesalts or esters thereof. Metformin (Glucophage®) is indicated forpatients with non-insulin dependent diabetes mellitus, particularlythose with refractory obesity. Physician's Desk Reference® page1080-1086, (56th ed. 2002).

Protein tyrosine phosphatase-1B (PTP-1B) inhibitors useful in thepresent disclosure include, but are not limited to, A-401,674, KR 61639,OC-060062, OC-83839, OC-297962, MC52445, MC52453, and the compoundsdisclosed in WO 02/26707, WO 02/26743, JP 2002114768, andpharmaceutically acceptable salts or esters thereof.

Dipeptidyl peptidase IV (DPP-IV) inhibitors, such as isoleucinethiazolidide; NVP-DPP728; P32/98; and LAP 237, P 3298, TSL 225, valinepyrrolidide, TMC-2A/2B/2C, CD-26 inhibitors, FE 999011, P9310/K364, VIP0177, DPP4, SDZ 274A444; and the compounds disclosed in WO 03/00449; WO03/004496; EP 1 258 476; WO 02/083128; WO 021062764; WO 03/000250; WO03/002530; WO 03/002531; WO 03/002553; WO 03/002593; WO 03/000180; andWO 03/000181.

Sulfonylureas useful in the present disclosure include, but are notlimited to, acetohexamide, chlorpropamide, diabinese, glibenclamide,glipizide, glyburide, glimepiride, gliclazide, glipentide, gliquidone,glisolamide, tolazamide, and tolbutamide, pharmaceutically acceptablesalts or esters thereof. Meglitinides useful in the present disclosureinclude, but are not limited to, repaglinide and nateglinide, andpharmaceutically acceptable salts or esters thereof.

Alpha glucoside hydrolase inhibitors (or glucoside inhibitors) useful inthe present disclosure include, but are not limited to, acarbose,adiposine, camiglibose, emiglitate, miglitol, voglibose, pradimicin-Q,salbostatin, CKD-711, MDL-25,637, MDL-73,945, and MOR 14, andpharmaceutically acceptable salts or esters thereof, and the compoundsdisclosed in U.S. Pat. Nos. 4,062,950, 4,174,439, 4,254,256, 4,701,559,4,639,436, 5,192,772, 4,634,765, 5,157,116, 5,504,078, 5,091,418,5,217,877, and 5,091,524. Alpha-amylase inhibitors useful in the presentdisclosure include, but are not limited to, tendamistat, trestatin, andA1-3688, and pharmaceutically acceptable salts and esters thereof, andthe compounds disclosed in U.S. Pat. Nos. 4,451,455, 4,623,714, and4,273,765.

Insulin secreatagogues useful in the present disclosure include, but arenot limited to, linogliride and A-4166, and pharmaceutically acceptablesalts and esters thereof.

Fatty acid oxidation inhibitors useful in the present disclosureinclude, but are not limited to, clomoxir, and etomoxir, andpharmaceutically acceptable salts and esters thereof. A2 antagonistsuseful in the present disclosure include, but are not ‘limited to,midaglizole, isaglidole, deriglidole, idazoxan, earoxan, fluparoxan, andpharmaceutically acceptable salts and esters thereof. Insulin or insulinmimetics useful in the present disclosure include, but are not limitedto, biota, LP-100, novarapid, insulin detemir, insulin lispro, insulinglargine, insulin zinc suspension (lente and ultralente), Lys-Proinsulin, GLP-1 (73-7) (insulintropin), and GLP-1 (7-36)-NH2), andpharmaceutically acceptable salts or esters thereof.

Glycogen phosphorylase inhibitors useful in the present disclosureinclude, but are not limited to, CP-368, 296, CP-316,819, BAYR3401, andcompounds disclosed in WO 01/94300, and WO 02/20530, andpharmaceutically acceptable salts or esters thereof. GLP-1 agonistsuseful in the present disclosure include, but are not limited to,exendin-3 and exendin-4, and compounds disclosed in US 2003087821 and NZ504256, and pharmaceutically acceptable salts or esters thereof.

Non-thiazolidinediones useful in the present disclosure include, but arenot limited to, JT-501, and farglitazar (GW-2570/GI-262579), andpharmaceutically acceptable salts or esters thereof. Glycokinaseactivators useful in this disclosure, include, but are not limited to,fused heteroaromatic compounds such as those disclosed in US 2002103199,and isoindolin-1-one-substituted propionamide compounds such as thosedisclosed in WO 02/48106.

Serotonin (5HT) transport inhibitors useful in this disclosure include,but are not limited to, paroxetine, fluoxetine, fenfluramine,fluvoxamine, sertraline, and imipramine Norepinephrine (NE) transportinhibitors useful in this disclosure include, but are not limited to, GW320659, despiramine, talsupram, and nomifensine. Cannabinoid receptor 1(CB-1) antagonist/inverse agonists useful in the present disclosureinclude: U.S. Pat. Nos. 5,532,237, 4,973,587, 5,013,837, 5,081,122,5,112,820, 5,292,736, 5,624,941 and U.S. Pat. No. 6,028,084, and PCTApplication Nos. WO 96/33159, WO 98/33765, WO98/43636, WO98/43635, WO01/09120, WO 98/31227, WO 98/41519, WO 98/37061, WO 00/10967, WO00/10968, WO 97/29079, WO 99/02499, WO 01/58869, WO 02/076949, WO01/64632, WO 01/64633, WO 01/64634, and WO 03/007887, and EPOApplication No. EP-658546. Specific CB-1 antagonists/inverse agonistsuseful in the present disclosure include, but are not limited to,rimonabant (Sanofi Synthelabo), SR-147778 (Sanofi Synthelabo), BAY65-2520 (Bayer), and SLY 319 (Solvay). CCK-A agonists useful in thepresent disclosure include GI 181771, and SR 146,131. Ghrelinantagonists useful in the present disclosure, include: PCT ApplicationNos. WO 01/87335, and WO 02/08250. Histamine 3 (H3) antagonist/inverseagonists useful in the present disclosure include: PCT Application No.WO 02/15905, and O-[3-(1H-imidazol4-yl)propanol]carbamates(Kiec-Kononowicz, K. et al., Pharmazie, 55:349-55 (2000)),piperidine-containing histamine H3-receptor antagonists (Lazewska, D. etal., Pharmazie, 56:927-32 (2001), benzophenone derivatives and relatedcompounds (Sasse, A. et al. Arch. Pharm. (Weinheim) 334:45-52 (2001)),substituted N-phenyl carbamates (Reidemeister, S. et al., Pharmazie,55:83-6 (2000)), and proxifan derivatives (Sasse, A. et al., J. Med.Chem. 43:3335-43 (2000)). Specific H3 antagonists/inverse agonistsuseful in the present disclosure include, but are not limited to,thioperamide, 3-(1H-imidazol4-yl)propyl N-4-pentenyl)carbamate,clobenpropit, iodophenpropit, imoproxifan, GT2394 (Gliatech), andA331440.

Melanin-concentrating hormone receptor (MCHLR) antagonists andmelanin-concentrating hormone 2 receptor (MCH2R) agonist/antagonistsuseful in the present disclosure include PCT Patent Application Nos. WO01/82925, WO 01/87834, WO 02/06245, WO 02/04433, and WO 02/51809, andJapanese Patent Application No. JP 13226269. Specific MCH1R antagonistsuseful in the present disclosure include, but are not limited to,T-226296 (Takeda), SB 568849, and SNAP 7941. Neuropeptide Y1 (NPY1)antagonists useful in the present disclosure, include: U.S. Pat. No.6,001,836, and PCT Application Nos. WO 96/14307, WO 01/23387, WO99/51600, WO 01/85690, WO 01/85098, WO 01/85173, and WO 01/89528.Specific examples of NPY1 antagonists useful in the present disclosureinclude, but are not limited to, BIBP3226, J-115814, BIBO 3304,LY-357897, CP-671906, and GI-264879A. Neuropeptide Y2 (NPY2) agonistsuseful in the present disclosure, include, but are not limited to,peptide YY (PYY), and PYY3_36, peptide YY analogs, PYY agonists, and thecompounds disclosed in WO 03/026591, WO 03/057235, and WO 03/027637.Neuropeptide Y5 (NPY5) antagonists useful in the present disclosure,include, but are not limited to, the compounds described in: U.S. Pat.Nos. 6,140,354, 6,191,160, 6,258,837, 6,313,298, 6,337,332, 6,329,395,and 6,340,683, U.S. Pat. Nos. 6,326,375, 6,329,395, 6,337,332,6,335,345, European Patent Nos. EP-01010691, and EP 01044970, andPCT-International Patent Publication Nos. WO 97/19682, WO 97/20820, WO97/20821, WO 97/20822, WO 97/20823, WO 98/27063, WO 00/107409,WO00/185714, WO 00/185730, WO 00/64880, WO 00/68197, WO 00/69849, wo01/09120, wo 01/85714, WO 01/85730, WO 01/07409, WO 01/02379, WO01/02379, WO 01/23388, WO 01/23389, WO 01/44201, WO 01/62737, WO01/62738, WO 01/09120, WO 02/20488, WO 02/22592, WO 02/48152, WO02/49648, and WO 01/14376. Specific NPY5 antagonists useful in thecombinations of the present disclosure, include, but are not limited toGW-569180A, GW-594884A, GW-587081X, GW-548118X, FR 235,208, FR226928, FR240662, FR252384, 1229U91, GI-264879A, CGP71683A, LY-377897, LY366377,PD-160170, SR-120562A, SR-120819A, JCF-104, and H409/22. Additionalspecific NPY5 antagonists useful in the combinations of the presentdisclosure, include, but are not limited to the compounds described inNorman et al., J. Med. Chem. 43:42884312 (2000). Leptin includes, but isnot limited to, recombinant human leptin (PEG-OB, Hoffman La Roche) andrecombinant methionyl human leptin (Amgen). Leptin derivatives (e.g.,truncated forms of leptin) useful in the present disclosure include:U.S. Pat. Nos. 5,552,524, 5,552,523, 5,552,522, 5,521,283, and PCTInternational Publication Nos. WO 96/23513, WO 96/23514, WO 96/23515, WO96/23516, WO 96/23517, WO 96/23518, WO 96/23519, and WO 96/23520.

Opioid antagonists useful in the present disclosure include: PCTApplication No. WO 00/21509. Specific opioid antagonists useful in thepresent disclosure include, but are not limited to, nalmefene (Revex®),3-methoxynaltrexone, naloxone, and naltrexone. Orexin antagonists usefulin the present disclosure include: PCT Patent Application Nos. WO01/96302, WO 01/68609, WO 02/51232, WO 02/51838, and WO 03/023561.Specific orexin antagonists useful in the present disclosure include,but are not limited to, SB-334867-A. Acyl-estrogens useful in thepresent disclosure include oleoyl-estrone (del Mar-Grasa, M. et al.,Obesity Research, 9:202-9 (2001)). Cholecystokinin-A (CCK-A) agonistsuseful in the present disclosure include U.S. Pat. No. 5,739,106.Specific CCK-A agonists include, but are not limited to, AR-R 15849,GI181771, JMv-180, A-71378, A-71623 and SR146131. Specific ciliaryneurotrophic factors (CNTh) useful in the present disclosure include,but are not limited to, GI-181771 (Glaxo-SmithKline), SR146131 (SanofiSynthelabo), butabindide, PD170,292, PD 149164 (Pfizer). CNTFderivatives useful in the present disclosure include, but are notlimited to, axokine (Regeneron), and PCT Application Nos. WO 94/09134,WO 98/22128, and WO 99/43813. Growth hormone secretagogue (GHS) agonistsuseful in the present disclosure include: U.S. Pat. No. 6,358,951, andU.S. Patent Application Nos. 2002/049196 and 2002/022637, and PCTApplication Nos. WO 01/56592, and WO 02/32888. Specific GHS agonistsinclude, but are not limited to, NN703, hexarelin, MK-0677, SM-130686,CP424 391, L-692,429 and L-163,255.

5HT2c agonists useful in the present disclosure include: U.S. Pat. No.3,914,250, and PCT Application Nos. WO 02/36596, WO 02/48124, WO02/10169, WO 01/66548, WO 02/44152, WO 02/51844, WO 02/40456, and WO02/40457. Specific 5HT2c agonists useful in this disclosure include, butare not limited to, BVT933, DPCA37215, 1K264, PNU 22394, WAY161503,R-1065, and YM 348.

Mc4r agonists useful in the present disclosure include: PCT ApplicationNos. WO 99/64002, WO 00/74679, WO 01/991752, WO 01/74844, WO 01/70708,WO 01/70337, WO 01/91752, WO 02/059095, WO 02/059107, WO 02/059108, WO02/059117, wo 02/12166, WO 02111715, WO 02/12178, WO 02/15909, WO02/068387, WO 02/068388, WO 02/067869, WO 03/007949, and WO 03/009847.Specific Mc4r agonists useful in the present disclosure include CIR86036(Chiron), ME-10142, and ME-10145 (Melacure).

Monoamine reuptake inhibitors useful in the present disclosure include:PCT Application Nos. WO 01/27068, and WO 01/62341. Specific monoaminereuptake inhibitors useful in the present disclosure include, but arenot limited to, sibutramine (Meridia O/Reductil®) disclosed in U.S. Pat.Nos. 4,746,680, 4,806,570, and 5,436,272, and U.S. Patent PublicationNo. 2002/0006964.

Serotonin reuptake inhibitors, and releasers, useful in the presentdisclosure include: dexfenfluramine, fluoxetine, and other serotoninreuptake inhibitors, including, but not limited to, those in U.S. Pat.No. 6,365,633, and PCT Patent Application Nos. WO 01/27060, and WO01/162341.

11 β HSD-1 inhibitor useful in the present disclosure include, but arenot limited to, BVT 3498, BVT 2733, and those compounds disclosed in WO01/90091, WO 01/90090, WO 01/90092. Uncoupling Protein (UCP-1, UCP-2,and UCP-3) activators useful in the present disclosure include: PCTPatent Application No. WO 99/00123. Specific uncoupling protein (UCP-1,UCP-2, and UCP-3) activators useful in the present disclosure include,but are not limited to, phytanic acid,4-[(E)-2-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-napthalenyl)-1-propenyl]benzoicacid (TTNPB), and retinoic acid.

β3 adrenergic receptor (β3) agonists useful in the present disclosureinclude: U.S. Pat. Nos. 5,705,515 and 5,451,677 and PCT PatentApplication Nos. WO 01/74782, and WO 02/32897. Specific β agonistsuseful in the present disclosure include, but are not limited to,AD9677/TAK677 (Dainippon/Takeda), CL-316,243, SB 418790, BRL-37344,L-796568, BMS-196085, BRL-35135A, CGP12177A, BTA-243, GW 427353,Trecadrine, Zeneca D7114, and SR 59119A.

Thyroid hormone β agonists useful in the present disclosure include: PCTApplication No. WO 02/15845 and Japanese Patent Application No. JP2000256190. Specific thyroid hormone β agonists useful in the presentdisclosure include, but are not limited to, KB-2611 (KaroBioBMS).Specific fatty acid synthase (PAS) inhibitors useful in the presentdisclosure, include, but are not limited to, Cerulenin and C75. Specificphosphodieterase (PDE) inhibitors useful in the present disclosure,include, but are not limited to, theophylline, pentoxifYlline,zaprinast, sildenafil, arnrinone, milrinone, cilostamide, rolipram, andcilomilast.

Lipase inhibitors useful in the present disclosure include, but are notlimited to, those disclosed in PCT Application No. WO 01/77094, and U.S.Pat. Nos. 4,598,089, 4,452,813, 5,512,565, 5,391,571, 5,602,151,4,405,644, 4,189,438, and 4,242,453. Specific lipase inhibitors usefulin the present disclosure include, but are not limited to,tetrahydrolipstatin (orlistat/Xenical®), Triton WR1339, RHC80267,lipstatin, teasaponin, and diethylumbelliferyl phosphate, FL-386,WAY-121898, Bay-N-3176, valilactone, esteracin, ebelactone A, ebelactoneB, and RHC 80267.

Examples of HMG-CoA reductase inhibitors include, but are not limitedto, lovastatin, simvastatin, pravastatin and fluvastatin. Examples ofHMG-CoA synthase inhibitors are the beta-lactone derivatives disclosedin U.S. Pat. Nos. 4,806,564, 4,816,477, 4,847,271, and 4,751,237; thebeta-lactam derivatives disclosed in U.S. Pat. No. 4,983,597 and U.S.Ser. No. 07/540,992 tiled Jun. 20, 1990; and the substitutedoxacyclopropane analogues disclosed in European Patent Publication EP 0411 703. Examples of squalene epoxidase inhibitors are disclosed inEuropean Patent Publication EP 0 318 860 and in Japanese PatentPublication J02 169-571A. Examples of LDL-receptor gene inducermolecules are disclosed in U.S. Pat. No. 5,182,298 filed Mar. 18, 1991.Other cholesterol lowering agents that may be administered includeniacin, probucol, fibric acids (i.e., clofibrate and gemfibrozil), andLDL-receptor gene inducers.

Examples of PARP inhibitors include, but are not limited to,iodonitocoumarin, 5-iodo-6-nitrocournarin,3,4-dihydro-5-methyl-isoquinolinone 4-amino-1,8-naphthalimide,3-methoxybenzamide, 8-hydroxy-2-methyl-3-hydro-quinazolin-4-one,2-{3-[4-(4-fluorophenyl)-3,6-dihydro-1(2h)-pyridinyl]propyl}-8-methyl-4(3h)-quinazolinone,5-fluoro-1-[4-(4-phenyl-3,6-dihydropyridin-1(butyl]quinazoline-2,4(1h,3h)-dione,3-(4-chlorophenyl) quinoxaline-5-carboxamide,2-(3′-methoxyphenyl)benzirnidazole-4-carboxam, benzamide,3-aminobenzamide, 3-aminophtalhydrazide, and 1,5-dihydroxyisoquinoline.

The above-mentioned compounds, which can be used in combination with acompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof, can beprepared and administered as described in the art such as in thedocuments cited above.

The above compounds are only illustrative of the ACMSD inhibitors,anti-diabetic agents, anti-obesity agents, cholesterol lower agent,compounds that boost NAD⁺ levels, compounds that inhibit NAD⁺consumption that can be used in the compositions of the presentdisclosure. As this listing of compounds is not meant to becomprehensive, the methods of the present disclosure may employ anyanti-obesity agent and any anti-diabetic agent, and are not limited toany particular structural class of compounds.

As used herein, “combination therapy” includes the administration of acompound of the present disclosure, or a pharmaceutically acceptablesalt, prodrug, metabolite, polymorph or solvate thereof, and at least asecond agent as part of a specific treatment regimen intended to providethe beneficial effect from the co-action of these therapeutic agents.The beneficial effect of the combination includes, but is not limitedto, a cooperative, e.g., synergistic, effect and/or a pharmacokinetic orpharmacodynamic co-action, or any combination thereof, resulting fromthe combination of therapeutic agents. Administration of thesetherapeutic agents in combination typically is carried out over adefined time period (usually minutes, hours, days or weeks dependingupon the combination selected). “Combination therapy” may be, butgenerally is not, intended to encompass the administration of two ormore of these therapeutic agents as part of separate monotherapyregimens that incidentally and arbitrarily result in the combinations ofthe present disclosure.

“Combination therapy” is intended to embrace administration of thesetherapeutic agents in a sequential manner, wherein each therapeuticagent is administered at a different time and in any order, or inalternation and in any order, as well as administration of thesetherapeutic agents, or at least two of the therapeutic agents, in asubstantially simultaneous manner. Substantially simultaneousadministration can be accomplished, for example, by administering to thesubject a single capsule having a fixed ratio of each therapeutic agentor in multiple, single capsules for each of the therapeutic agents.Sequential or substantially simultaneous administration of eachtherapeutic agent can be effected by any appropriate route including,but not limited to, oral routes, intravenous routes, intramuscularroutes, and direct absorption through mucous membrane tissues. Thetherapeutic agents can be administered by the same route or by differentroutes. For example, a first therapeutic agent of the combinationselected may be administered by intravenous injection while the othertherapeutic agents of the combination may be administered orally.Alternatively, for example, all therapeutic agents may be administeredorally or all therapeutic agents may be administered by intravenousinjection. The sequence in which the therapeutic agents are administeredis not narrowly critical.

All percentages and ratios used herein, unless otherwise indicated, areby weight. Other features and advantages of the present disclosure willbecome apparent from the different examples. The provided examplesillustrate different components and methodology useful in practicing thepresent disclosure. Generally speaking, the disclosure extends to anynovel one, or any novel combination, of the features disclosed in thisspecification (including the accompanying claims and drawings). Theexamples do not limit the claimed disclosure. Thus, features, integers,characteristics, compounds or chemical moieties described in conjunctionwith a particular aspect, embodiment or example of the disclosure are tobe understood to be applicable to any other aspect, embodiment orexample described herein, unless incompatible therewith. Based on thepresent disclosure the skilled artisan can identify and employ othercomponents and methodology useful for practicing the present disclosure.Moreover, unless stated otherwise, any feature disclosed herein may bereplaced by an alternative feature serving the same or a similarpurpose.

The Disclosure will now be described by way of example only withreference to the Examples below:

EXEMPLIFICATION

I. Compound Preparation

General Methods and Materials

All chemicals were purchased from Sigma-Aldrich, Alfa Aesar. ¹H NMRspectra were recorded at 200 and 400 MHz and ¹³C NMR spectra wererecorded at 100.6 and 50.3 MHz by using deuterated solvents indicatedbelow. TLC were performed on aluminium backed silica plates (silica gel60 F254). All the reactions were performed under nitrogen atmosphereusing distilled solvents. All tested compounds were found to have >95%purity determined by HPLC analysis. HPLC-grade water was obtained from atandem Milli-Ro/Milli-Q apparatus. The analytical HPLC measurements weremade on a Shimadzu LC-20AProminence equipped with a CBM-20Acommunication bus module, two LC-20AD dual piston pumps, a SPD-M20Aphotodiode array detector and a Rheodyne 7725i injector with a 20 μLstainless steel loop.

Example 1 Preparation of Intermediate 1.4

To a stirred solution of compound 1.1 (0.52 ml, 4.9 mmol), 1.2 (372 mg,4.9 mmol) and 1.3 (0.5 mL, 0.83 mL, 4.9 mmol) in ethanol (25 mL) wasadded K₂CO₃ (812 mg, 5.88 mmol). Stirring was continued at refluxovernight. The pale yellow solid was collected after cooling, taken upwith boiling water and filtered again. The aqueous phase was acidifiedto pH 5 with AcOH (15 drops), the precipitate was filtered and driedunder vacuum. The title compound 1.4 was obtained as pale yellow solid(500 g, 2.18 mmol). Yield 44%.

Example 2 Preparation of Intermediate 2.2

To a stirred solution of compound 1.1 (0.96 g, 8.8 mmol), 1.2 (672 mg,8.8 mmol) and 2.1 (1 g, 0.83 mL) in ethanol (55 mL) was added K₂CO₃(1.57 g, 11.44 mmol). Stirring was continued at reflux overnight. Theyellowish solid was collected after cooling, taken up with hot water andfiltered again. The aqueous phase was acidified to pH 1, the precipitatewas filtered and dried under vacuum. The title compound 2.2 was obtainedas yellowish solid (1 g, 4.25 mmol). Yield 49%. ¹H NMR (200 MHz, DMSO) δ7.22 (m, 1H), 7.68 (m, 1H), 7.85 (d, J=4.8 Hz, 1H), 8.05 (s, 1H).

Example 3 Preparation of Intermediate 3.2

To a stirred solution of compound 1.1 (0.96 mL, 8.8 mmol), 1.2 (672 mg,8.8 mmol) and 3.1 (1 g, 1.29 mL) in ethanol (55 mL) was added K₂CO₃(1.57 g, 11.44 mmol). Stirring was continued at reflux overnight. Theyellowish solid was collected after cooling, taken up with hot water andfiltered again. The aqueous phase was acidified to pH 1, the precipitatewas filtered and dried under vacuum. The title compound 3.2 was obtainedas yellowish solid (1 g, 4.25 mmol). Yield 49%.

Example 4 Preparation of Intermediate 4.2

To a stirred solution of compound 1.1 (1.42 mL, 13.37 mmol), 1.2 (1.01g, 13.3 mmol) and 4.1 (1.62 mL, 13.3 mL) in ethanol (50 mL) was addedpiperidine (2.64 mL, 26.7 mmol). Stirring was continued at refluxovernight. The solid was collected after cooling, taken up with hotwater and filtered again. The aqueous phase was acidified to pH1 andextracted with EtOAc (3×25 mL). The organic phase was washed with brineand dried over Na₂SO₄. The crude of reaction was purified by flashchromatography (CHCl₃/MeOH as gradient, from 0 to 2% for product),affording the title compound 4.2 (930 mg, 3.95 mmol) as white solid.Yield 30%.

Example 5 Preparation of Intermediate 5.2

To a stirred solution of compound 1.1 (0.49 mL, 4.67 mmol), 1.2 (355 mg,4.67 mmol) and 5.1 (0.44 mL, 4.67 mmol) in ethanol (25 mL) was addedK₂CO₃ (773 mg, 5.6 mmol). Stirring was continued at reflux overnight.The white solid was collected after cooling, dried under vacuum and usedfor the next step without further purification. The title compound 5.2was obtained as white solid (300 mg, 1.3 mmol). Yield 29%. ¹H NMR (400MHz, DMSO) δ 7.64 (d, J=4.7 Hz, 2H), 8.78 (d, J=4.7 Hz, 2H), 12.98 (s,1H).

Example 6 Preparation of Intermediate 6.2

To a stirred solution NaOEt (1.02 mL, 2.73 mmol) in EtOH abs (20 mL) wasadded compound 6.1 (500 mg, 2.73 mmol) and 1.2 (207 mg, 2.73 mmol).Stirring was continued at reflux 4 h. The volatiles were removed undervacuum. The crude of reaction was taken up with water and acidified withAcOH. The precipitate was collected dissolved in water, washed with amixture of CHCl₃ and MeOH. The aqueous phase was extracted with EtOAc(3×20 mL). The collected organic phase was washed with brine, dried overNa₂SO₄. The title compound 6.2 was obtained as white solid (250 mg, 1.49mmol). Yield 55%.

Example 7a Preparation of Intermediate 7.2

To a stirred solution of compound 1.1 (0.14 mL, 1.3 mmol) and 7.1 (150mg, 1.3 mmol) in EtOH (5 mL) was added piperidine (1 drop). Stirring wascontinued at room temperature overnight. The solvent was removed undervacuum. The crude of reaction was purified by flash chromatographyaffording the title compound 7.2 (160 mg, 0.77 mmol) as yellowish solid.Yield 58%.

Example 7b Preparation of Intermediate 7.3

To a stirred suspension of compound 7.2 (150 mg, 0.72 mmol) and compound1.2 (55 mg, 0.72 mmol) in EtOH (5 mL) was added K₂CO₃ (99 mg, 0.72mmol). Stirring was continued at reflux overnight. The white precipitatewas collected and used as well for the next step without furtherpurification. The title compound 7.3 (150 mg, 0.48 mmol) was obtained asyellowish solid as di-potassium salt. Yield 67%.

Example 8a Preparation of Intermediate 8.2

To a stirred solution of NH₂OH*HCl and NaHCO₃ in water (7 mL) wasgradually added a solution of m-tolunitrile (8.1) (2 mL, 17.0 mmol) inEtOH (13.3 mL). Stirring was continued at 80° C. for 4 h. The volatileswere removed under vacuum. The crude of reaction was taken up withwater, extracted with EtOAc (3×25 mL). The organic phase were collected,washed with brine and dried over Na₂SO₄ affording the title compound 8.2(1.5 g, 9 mmol) as white solid. Yield 59%.

Example 8b Preparation of Intermediate 8.3

To a solution of compound 8.2 (1 g, 6 mmol) in dry acetone (5 mL), wasadded dropwise at 0° C. EtOCOCl (0.63 mL, 6.6 mmol). Stirring wascontinued at this temperature for 1 h. Then a 5% NaOH solution was addedto the mixture. Stirring was continued for additional 1 h. The solventws removed under vacuum. The crude of reaction was poured in water,extracted with EtOAc (3×50 mL). The collected organic phase was washedwith brine, dried over Na₂SO₄. The title compound 8.3 (600 mg, 2.7 mmol)was obtained as white solid. Yield 45%.

Example 8c Preparation of Intermediate 8.4

To a solution of compound 8.3 (300 mg, 1.35 mmol) in EtOH abs (5 mL) wasadded sodium (50 mg) portion wise. Stirring was continued at roomtemperature for additional 4 h. The reaction was quenched by theaddition of MeOH. The solvent was removed under reduced pressure and thecrude was purified by flash chromatography. The title compound 8.4 (150mg, 0.85 mmol) was obtained as white solid. Yield 63%.

Example 8d Preparation of Intermediate 8.5

To a suspension of compound 8.4 (326 mg, 1.85 mmol) in CCl₄ (10 mL) wasadded AIBN (60.7 mg, 0.37 mmol) and NBS (493 mg, 2.77 mmol). Stirringwas continued at reflux overnight. The solvent was removed under reducedpressure. The reaction was taken up with water, extracted with EtOAc(3×20 mL) washed with brine and dried over Na₂SO₄. The crude waspurified by flash chromatography, eluting with Petroleum ether (Pet.Ether)/EtOAc (30% for product) affording the title compound 8.5 (280 mg,1.09 mmol) was obtained as white solid. Yield 59%.

Example 9 Preparation of Intermediate 9.2

To a suspension of compound 9.1 (750 mg, 5 mmol) in CCl₄ (15 mL) wasadded AIBN (41 mg, 0.25 mmol) and NBS (933.7 mg, 5.24 mmol). Stirringwas continued at reflux overnight. The solvent was removed under reducedpressure. The reaction was taken up with water, extracted with EtOAc(3×20 mL) washed with brine and dried over Na₂SO₄. The crude waspurified by flash chromatography, eluting with CH₂Cl₂/MeOH (3% forproduct) affording the title compound 9.2 (800 mg, 3.49 mmol) as whitesolid. Yield 70%.

Example 10a Preparation of Intermediate 10.2

A mixture of compound 10.1 (1.02 mL, 8.54 mmol), NaN₃ (832 mg, 12.8mmol) and Et₃N.HCl (1.76 g, 12.8 mmol) was heated at reflux 4 h. Thesolvent was removed under vacuum. The crude was poured in water,acidified to pH 1 with 3N HCl and extracted with EtOAc (3×20 mL). Theorganic phase was washed with brine, dried over Na₂SO₄ and concentratedunder reduced pressure. The title compound 10.2 (1.22 g, 7.6 mmol) wasobtained as white solid. Yield 89%.

Example 10b Preparation of Intermediate 10.3

To a suspension of compound 10.2 (300 mg, 1.87 mmol) in CH₃CN (15 mL)was added AIBN (31 mg, 0.18 mmol) and NBS (333 mg, 1.87 mmol). Stirringwas continued at reflux overnight. The solvent was removed under reducedpressure. The reaction was taken up with water, extracted with EtOAc(3×20 mL) washed with brine and dried over Na₂SO₄. The crude waspurified by flash chromatography, eluting with CH₂Cl₂/MeOH (7% forproduct) affording the title compound 10.3 (150 mg, 0.62 mmol) as lightyellow solid. Yield 34%.

Example 11 Preparation of Intermediate 11.3

To a solution of compound 11.1 (2.5 g, 23 mmol) in CH₂Cl₂ (25 mL) wasadded pyridine (1.63 mL, 20.3 mmol) and compound 11.2 (1.68 mL, 20.3mmol). Stirring was continued at room temperature overnight. The solventwas removed under reduced pressure. The reaction was taken up withwater, extracted with CH₂Cl₂ (3×30 mL) washed with brine and dried overNa₂SO₄. The crude was purified by flash chromatography, eluting withPet. Ether/EtOAc (25% for product) affording the title compound 11.3(735 mg, 3.19 mmol) as brownish solid. Yield 14%.

Example 12 Preparation of Intermediate 12.2

To a solution of compound 12.1 (2 g, 10.41 mmol) in EtOH (15 mL) wasadded EtONa (7 mL, 18.7 mmol) and compound 1.2 (1.18 g, 15.61 mmol).Stirring was continued at reflux overnight. The solvent was removedunder reduced pressure. The reaction was taken up with water, acidifiedto pH 3, extracted with EtOAc (3×20 mL) washed with brine and dried overNa₂SO₄. The crude was purified by flash chromatography, eluting withCH₂Cl₂/MeOH (2.5% for product) affording the title compound 12.2 (500mg, 2.44 mmol) as white solid. Yield 24%.

Example 13a Preparation of Intermediate 13.2

To a stirred solution of DIPA (7.6 mL, 54 mmol) in THF (53 mL) was addedn-BuLi (21.6 mL) at 0° C. Stirring was continued at this temperature 10minutes. The mixture was then cooled to −78° C. and EtOAc (2.4 mL, 27mmol) was added dropwise. Stirring was continued at this temperature 30minutes. After that, a solution of compound 13.1 (3 mL, 27 mmol) in THF(20 mL) was added dropwise. The reaction was allowed to warm to roomtemperature and was stirred overnight. The crude of reaction was pouredin water and extracted with EtOAc (3×30 mL). The collected organic phasewere washed with brine, dried over Na₂SO₄ and concentrated under vacuum.The title compound 13.2 was obtained as brownish oil (4.8 g, 24.3 mmol).Yield 90%.

Example 13b Preparation of Intermediate 13.3

To a solution of intermediate 13.2 (2 g, 10 mmol) in EtOH (15 mL) wasadded EtONa (21% wt/wt in EtOH) (7.5 mL, 20 mmol) and compound 1.2 (1.15g, 15.1 mmol). Stirring was continued at reflux overnight. The solventwas removed under reduced pressure. The reaction was taken up withwater. At pH 10 was recovered unreacted starting material. The mixturewas then acidified to pH 5, extracted with EtOAc (3×20 mL) washed withbrine and dried over Na₂SO₄. The crude was purified by flashchromatography, eluting with CH₂Cl₂/MeOH (7% for product) affording thetitle compound 13.3 (435 mg, 2.06 mmol) as yellowish solid. Yield 21%.

Example 14 Preparation of Compound 1

To a stirred suspension of intermediate 1.4 (1.6 g, 6.98 mmol) and K₂CO₃(2.88 g, 20.9 mmol) in CH₃CN (80 mL) was added 3-(chloromethyl)benzoicacid (1.19 g, 6.98 mmol). Stirring was continued overnight at reflux.The volatiles were removed under vacuo. The crude was taken up withwater, acidified to pH 5 and washed with EtOAc to remove impurities.Then the pH was adjusted to 3/4 and the mixture was extracted with EtOAc(3×50 mL). Titration with hot acetone afforded compound 1 (936 mg, 2.78mmol) as yellowish solid. Yield 40%. ¹H NMR (400 MHz, DMSO) δ 4.58 (s,2H), 7.44 (t, J=7.5 Hz, 1H), 7.54-7.61 (m, 3H), 7.67 (d, J=7.1 Hz, 1H),7.83 (d, J=7.5 Hz, 1H), 7.91 (d, J=7.27 Hz, 2H), 8.04 (s, 1H), 13 (s,1H); ¹³C NMR (100 MHz, DMSO) δ 33.5, 93.2, 115.6, 128.2, 128.4, 128.4,128.5, 128.5, 128.6, 129.7, 130.8, 131.5, 133.3, 135.1, 137.4, 165.4,166.8, 167.3. HPLC: 96.3%

Example 15 Preparation of Compound 4

To a stirred suspension of intermediate 2.2 (250 mg, 1.06 mmol) andK₂CO₃ (440 mg, 3.18 mmol) in CH₃CN (15 mL) was added3-(chloromethyl)benzoic acid (180 mg, 1.06 mmol). Stirring was continuedovernight at reflux. The volatiles were removed under vacuo. The crudewas taken up with water, washed with EtOAc, acidified to pH 1 andextracted with EtOAc (3×50 mL). Titration with hot acetone affordedcompound 4 (45 mg, 0.12 mmol) as yellowish solid. Yield 12%. ¹H NMR (400MHz, DMSO) δ 4.62 (s, 2H), 7.33 (t, J=4.3 Hz, 1H), 7.44 (t, J=7.6 Hz,1H), 7.72 (d, J=7.5 Hz, 1H), 7.82 (d, J=7.5 Hz, 1H), 8.05 (m, 2H), 8.26(d, J=3.8 Hz, 1H), 12.99 (s, 1H); ¹³C NMR (100 MHz, DMSO) δ 33.9, 88.7,116.5, 128.8, 129.3, 129.9, 130.2, 131.5, 132.1, 133.7, 135.4, 137.9,139.7, 159.0, 161.2, 165.3, 167.4. HPLC: 97.2%

Example 16 Preparation of Compound 3

To a stirred suspension of intermediate 3.2 (250 mg, 1.06 mmol) andK₂CO₃ (440 mg, 3.18 mmol) in CH₃CN (15 mL) was added3-(chloromethyl)benzoic acid (180 mg, 1.06 mmol). Stirring was continuedovernight at reflux. The volatiles were removed under vacuo. The crudewas taken up with water, washed with EtOAc, acidified to pH 1 andextracted with EtOAc (3×50 mL). Titration with a mixture of Et₂O/acetoneafforded compound 3 (260 mg, 0.7 mmol) as yellowish solid. Yield 70%. ¹HNMR (400 MHz, DMSO) δ 4.63 (s, 2H), 7.44 (t, J=7.6 Hz, 1H), 7.69 (d,J=7.7 Hz, 1H), 7.74 (dd, J=5 Hz, J=2.9 Hz, 1H), 7.84 (m, 2H), 8.05 (s,1H), 8.58 (m, 1H), 13.0 (s, 1H); ¹³C NMR (100 MHz, DMSO) δ 35.3, 90.1,118.0, 130.2, 130.7, 131.3, 131.6, 132.9, 133.5, 135.1, 136.8, 139.3,141.1, 160.4, 162.7, 166.7, 168.8. HPLC: 95.0%

Example 17 Preparation of Compound 6

To a stirred suspension of intermediate 4.2 (250 mg, 1.18 mmol) andK₂CO₃ (495 mg, 3.56 mmol) in CH₃CN (15 mL) was added3-(chloromethyl)benzoic acid (202 mg, 1.18 mmol). Stirring was continuedovernight at reflux. The volatiles were removed under vacuo. The crudewas taken up with water, washed with EtOAc, acidified to pH 1 andextracted with EtOAc (3×50 mL). Titration with Et₂O afforded compound 6(90 mg, 0.24 mmol) as white solid. Yield 21%. ¹H NMR (400 MHz, DMSO) δ1.24 (m, 3H), 1.60 (m, 7H), 2.74 (m, 1H), 4.52 (s, 2H), 7.45 (t, J=7.18Hz, 1H), 7.67 (d, J=6.83 Hz, 1H), 7.82 (d, J=7.17 Hz, 1H), 8.04 (s, 1H),13.0 (s, 1H). ¹³C NMR (100 MHz, DMSO) δ 25.4, 25.7, 25.7, 30.3, 30.3,33.8, 44.9, 94.1, 115.3, 128.6, 129.2, 130.1, 131.3, 133.6, 138.5,161.1, 166.2, 167.4, 177.9. HPLC: 98.1%

Example 18 Preparation of Compound 7

To a stirred suspension of intermediate 5.2 (220 mg, 0.95 mmol) andDIPEA (0.18 mL, 1.05 mmol) in DMSO (5 mL) was added3-(chloromethyl)benzoic acid (178 mg, 1.05 mmol). Stirring was continuedovernight at room temperature. The light yellow solid was collected,washed with crushed ice and water, and dried under vacuum. Triturationwith hot EtOAc afforded compound 7 (180 mg, 0.49 mmol) as light yellowsolid. Yield 53%. ¹H NMR (400 MHz, DMSO) δ 4.55 (s, 2H), 7.44 (m, 1H),7.66 (d, J=6 Hz, 1H), 7.81 (m, 3H), 8.02 (s, 1H), 8.80 (s, 2H), 13.1 (s,1H); ¹³C NMR (100 MHz, DMSO) δ 34.1, 94.8, 115.8, 122.8, 122.8, 128.7,129.2, 130.4, 131.3, 133.9, 138.1, 143.1, 150.5, 150.5, 161.8, 165.7,167.4, 167.4. HPLC: 95.1%

Example 19a Preparation of Compound 14

To a stirred suspension of intermediate 12.2 (100 mg, 0.43 mmol) andK₂CO₃ (178 mg, 1.29 mmol) in CH₃CN (15 mL) was added3-(chloromethyl)benzoic acid (74 mg, 0.43 mmol). Stirring was continuedovernight at reflux. The volatiles were removed under vacuo. The crudewas taken up with water, washed with EtOAc, acidified to pH 3 andextracted with EtOAc (3×50 mL). Titration with a mixture of Et₂O/Acetoneafforded compound 14 (30 mg, 0.088 mmol) as white solid. Yield 21%. ¹HNMR (400 MHz, DMSO) δ 4.59 (s, 2H), 6.69 (s, 1H), 7.41 (m, 1H), 7.46 (m,3H), 7.71 (d, J=7.5 Hz, 1H), 7.81 (d, J=7.74 Hz, 1H), 8.06 (m, 3H),12.85 (s, 2H). ¹³C NMR (100 MHz, DMSO) δ 33.8, 127.3, 127.3, 128.5,129.1, 129.2, 130.1, 131.0, 131.3, 131.5, 133.6, 136.3, 138.8, 167.5.

Example 19b Preparation of Compound 11

To a stirred solution of compound 14 (100 mg, 0.29 mmol) in acetic acid(5 mL) was added lead dioxide (77.2 mg, 0.32 mmol) and bromine (0.02 mL,0.32 mmol). Stirring was continued for 6 hrs at room temperature. Themixture was poured in a solution of Na₂S₂O₅ and was extracted with EtOAc(3×20 mL). The collected organic phases were washed with water andbrine, and then they were dried over Na₂SO₄. Titration with a mixture ofEt₂O/Acetone afforded compound 11 (40 mg, 0.09 mmol) as white solid.Yield 33%. ¹H NMR (400 MHz, DMSO) δ 4.44 (s, 2H), 7.42 (t, J=7.6 Hz,1H), 7.48 (m, 3H), 7.61-7.65 (m, 3H), 7.82 (d, J=7.5 Hz, 1H), 8.0 (s,1H), 13.1 (m, 2H). ¹³C NMR (100 MHz, DMSO) δ 33.9, 128.4, 128.6, 129.14,129.3, 129.3, 129.3, 130.1, 130.2, 131.3, 133.9, 138.1, 138.4, 167.5.HPLC: 94.2%

Example 20a Preparation of Compound 15

To a stirred suspension of intermediate 13.3 (235 mg, 1.11 mmol) andK₂CO₃ (460 mg, 3.33 mmol) in CH₃CN (15 mL) was added3-(chloromethyl)benzoic acid (190 mg, 1.11 mmol). Stirring was continuedovernight at reflux. The volatiles were removed under vacuo. The crudewas taken up with water, washed with EtOAc, acidified to pH 3 andextracted with EtOAc (3×50 mL). Titration with a mixture of Et₂O/Acetoneafforded compound 15 (150 mg, 0.44 mmol) as white solid. Yield 39%. ¹HNMR (400 MHz, DMSO) δ 4.55 (s, 2H), 6.64 (s, 1H), 7.19 (dd, J=4.9 Hz,J=3.8 Hz, 1H), 7.43 (t, J=7.7 Hz, 1H), 7.74 (d, J=7.6 Hz, 1H), 7.77 (d,J=4.9 Hz, 1H), 7.81 (d, J=7.8 Hz, 1H), 7.90 (d, J=3.6 Hz, 1H), 8.08 (s,1H), 12.80 (s, 2H). ¹³C NMR (100 MHz, DMSO) δ 33.5, 101.6, 128.1, 128.6,129.1, 129.1, 130.1, 131.1, 131.4, 133.7, 138.9, 141.7, 167.4.

Example 20b Preparation of Compound 12

To a stirred solution of compound 15 (134 mg, 0.39 mmol) in acetic acid(5 mL) was added lead dioxide (102 mg, 0.42 mmol) and bromine (0.022 mL,0.42 mmol). Stirring was continued for 6 hrs at room temperature. Themixture was poured in a solution of Na₂S₂O₅ and was extracted with EtOAc(3×20 mL). The collected organic phases were washed with water andbrine, and then they were dried over Na₂SO₄. The crude of reaction wassubjected to flash chromatography purification eluting with CH₂Cl₂/MeOH(10% for product). Compound 12 (45 mg, 0.11 mmol) was obtained as whitesolid. Yield 27%. ¹H NMR (400 MHz, DMSO) δ 4.56 (s, 2H), 7.27 (t, J=3.5Hz, 1H), 7.44 (t, J=7.6 Hz, 1H), 7.72 (d, J=7.5 Hz, 1H), 7.82 (d, J=7.7Hz, 1H), 7.92 (d, J=4.5 Hz), 8.07 (s, 1H), 8.33 (d, J=2.9 Hz, 1H), 13.1(m, 2H). ¹³C NMR (100 MHz, DMSO) δ 33.8, 128.6, 128.7, 128.7, 129.2,130.1, 131.4, 132.3, 132.8, 133.6, 138.3, 141.1, 152.1, 158.5, 159.6,167.4. HPLC: 95.2%

Example 21 Preparation of Compound 13

To a stirred solution of compound 14 (100 mg, 0.29 mmol) in acetic acid(5 mL) was added lead dioxide (55.8 mg, 0.35 mmol) andN-chlorosuccinimmide (47 mg, 0.35 mmol). Stirring was continued for 6hrs at room temperature. The mixture was poured in water and wasextracted with EtOAc (3×20 mL). The collected organic phases were washedwith water and brine, and then they were dried over Na₂SO₄. Titrationwith a mixture of Et₂O/acetone afforded compound 13 (40 mg, 0.1 mmol) aswhite solid. Yield 37%. ¹H NMR (400 MHz, DMSO) δ 4.47 (s, 2H), 7.43 (t,J=7.7 Hz, 1H), 7.49 (m, 3H), 7.64 (d, J=7.2 Hz, 1H), 7.71 (m, 2H), 7.83(d, J=7.5 Hz, 1H), 8.02 (s, 1H), 13.1 (s, 1H), 13.25 (s, 1H); ¹³C NMR(100 MHz, DMSO) δ 33.9, 128.4, 128.4, 128.6, 129.1, 129.4, 130.2, 131.3,131.3, 133.8, 136.5, 138.3, 167.5. HPLC: 95.3%

Example 22 Preparation of Compound 22

A stirred suspension of compound 1 (160 mg, 0.44 mmol) and POCl₃ (3 mL)was heated at 70° C. for 6 h. The white suspension turned red. Theexcess of POCl₃ was carefully destroyed with crushed ice and then water.The mixture was extracted with EtOAc (3×20 mL). The collected organicphase were washed with brine, dried over Na₂SO₄ and evaporated. Flashchromatography purification (gradient CH₂Cl₂/MeOH) afforded the titlecompound 22 (60 mg, 0.16 mmol) as white solid. Yield 36%. ¹H NMR (400MHz, DMSO) δ 4.58 (s, 2H), 7.44 (t, J=7.7 Hz, 1H), 7.58-7.62 (m, 2H),7.66 (d, J=7.17 Hz, 1H), 7.70 (d, J=7.7 Hz, 1H), 7.82 (d, J=7.7 Hz, 1H),7.94 (d, J=7.1 Hz, 2H), 8.08 (s, 1H), 12.98 (s, 1H); ¹³C NMR (100 MHz,DMSO) δ 34.9, 102.5, 115.2, 128.7, 129.2, 129.2, 129.6, 129.6, 130.4,131.4, 132.7, 133.9, 134.7, 138.0, 162.9, 167.5, 169.0, 174.3. HPLC:98.8%

Example 23 Preparation of Compound 10

To a stirred suspension of intermediate 6.2 (145 mg, 0.86 mmol) andK₂CO₃ (599 mg, 4.33 mmol) in CH₃CN (15 mL) was added3-(chloromethyl)benzoic acid (148 mg, 0.86 mmol). Stirring was continuedovernight at reflux. The volatiles were removed under vacuo. The crudewas taken up with water, washed with EtOAc, acidified to pH 3 andextracted with EtOAc (3×50 mL). The crude of reaction was purified byflash chromatography, eluting with (CH₂Cl₂/MeOH+ACOH 3%) affording thetitle compound 10 (60 mg, 0.2 mmol) as white solid. Yield 23%. ¹H NMR(400 MHz, DMSO) δ 2.44 (s, 3H), 4.49 (s, 2H), 7.45 (t, J=7.6 Hz, 1H),7.68 (d, J=7.4 Hz, 1H), 7.82 (d, J=7.6 Hz, 1H), 8.05 (m, 1H), 13.1 (s,1H); ¹³C NMR (100 MHz, DMSO) δ 23.3, 33.9, 95.3, 115.6, 128.7, 129.1,130.6, 131.2, 134.1, 138.0, 161.1, 165.7, 167.4, 170.9. HPLC 96.5%

Example 24 Preparation of Compound 5

To a stirred suspension of intermediate 7.3 (414 mg, 1.27 mmol) andK₂CO₃ (526 mg, 3.81 mmol) in CH₃CN (20 mL) was added3-(chloromethyl)benzoic acid (217 mg, 1.27 mmol). Stirring was continuedovernight at reflux. The volatiles were removed under vacuo. The crudewas taken up with water, washed with EtOAc, acidified to pH 3 andextracted with EtOAc (3×50 mL). The title compound 5 has been obtained(260 mg, 0.7 mmol) as pure light yellow solid after titration with amixture of Et₂O/Acetone. Yield 55%. ¹H NMR (400 MHz, DMSO) δ 4.62 (s,2H), 7.45 (m, 1H), 7.74 (d, J=5.9 Hz, 1H), 7.82 (d, J=6.1 Hz, 1H), 8.08(s, 1H), 8.21 (d, J=7.2 Hz, 2H), 12.9 (s, 1H); ¹³C NMR (100 MHz, DMSO) δ34.0, 90.5, 114.9, 127.7, 128.8, 129.3, 130.1, 131.4, 133.7, 138.1,146.2, 156.7, 161.9, 163.8, 166.5, 167.4. HPLC 96.5%

Example 25 Preparation of Compound 19

To a stirred suspension of intermediate 2.2 (100 mg, 0.42 mmol) andDIPEA (0.07 mL, 0.47 mmol) in DMSO (5 mL) was added intermediate 8.5(120 mg, 0.47 mmol). Stirring was continued overnight at roomtemperature. The crude was poured in water, washed with EtOAc thenacidified to pH 3 and extracted with EtOAc (3×50 mL). The title compound19 has been obtained (65 mg, 0.15 mmol) as pure orange solid after flashchromatography purification eluting with CH₂Cl₂/MeOH (10% for product).Yield 38%. ¹H NMR (400 MHz, DMSO) δ 4.37 (s, 2H), 7.20 (t, J=4 Hz, 1H),7.45 (t, J=7.6 Hz, 1H), 7.63 (t, J=7.2 Hz, 2H), 7.75 (d, J=4.3 Hz, 1H),7.88 (s, 1H), 8.07 (d, J=3 Hz, 1H); ¹³C NMR (100 MHz, DMSO) δ 33.7,85.7, 120.4, 124.9, 125.4, 126.7, 128.7, 128.8, 129.5, 131.1, 132.3,140.6, 142.2, 159.1, 159.9, 163.3, 170.4, 171.3. HPLC 94.1%.

Example 26 Preparation of Compound 18

To a stirred suspension of intermediate 2.2 (500 mg, 0. mmol) and DIPEA(0.4 mL, 2.12 mmol) in DMSO (5 mL) was added intermediate 9.2 (487 mg,2.12 mmol). Stirring was continued overnight at room temperature. Thecrude was poured in water, washed with EtOAc then acidified to pH 3 andextracted with EtOAc (3×50 mL). The title compound 18 has been obtained(200 mg, 0.52 mmol) as pure yellowish solid after flash chromatographypurification eluting with CH₂Cl₂/MeOH (10% for product) and titrationwith a mixture of Et₂O/Acetone. Yield 25%. ¹H NMR (400 MHz, DMSO) δ 3.49(s, 2H), 4.53 (s, 2H), 7.16 (d, J=6.8 Hz, 1H), 7.26 (t, J=7.2 Hz, 1H),7.36 (m, 3H), 8.05 (d, J=4.4 Hz, 1H), 8.27 (s, 1H), 12.13 (s, 1H); ¹³CNMR (100 MHz, DMSO) δ 34.3, 40.9, 88.5, 116.8, 127.6, 128.9, 129.1,129.8, 130.4, 131.9, 135.2, 135.8, 137.0, 139.9, 159.1, 161.6, 165.7,172.9. HPLC 95.8%.

Example 27 Preparation of Compound 17

To a stirred suspension of intermediate 2.2 (160 mg, 0.66 mmol) andDIPEA (0.09 mL, 0.55 mmol) in DMSO (3 mL) was added intermediate 10.3(171 mg, 0.55 mmol). Stirring was continued overnight at roomtemperature. The crude was poured in water, washed with EtOAc thenacidified to pH 3 and extracted with EtOAc (3×50 mL). The title compound17 has been obtained (90 mg, 0.22 mmol) as pure orange solid after flashchromatography purification eluting with CH₂Cl₂/MeOH (5% for product)and prior titration with a mixture of Et₂O/Acetone. Yield 23%. ¹H NMR(400 MHz, DMSO) δ 4.59 (s, 2H), 7.29 (t, J=4.6 Hz, 1H), 7.54 (t, J=7.6Hz, 1H), 7.68 (t, J=7.8 Hz, 1H), 7.91 (d, J=7.7 Hz, 1H), 7.96 (d, J=4.9Hz, 1H), 8.16 (s, 1H), 8.22 (d, J=3.8 Hz, 1H); ¹³C NMR (100 MHz, DMSO) δ33.9, 88.9, 117.6, 125.0, 126.2, 127.9, 129.6, 129.9, 131.2, 131.9,134.4, 139.3, 140.4, 155.8, 159.1, 163.8, 167.0. HPLC 96.2%.

Example 28 Preparation of Compound 23

To a stirred suspension of intermediate 2.2 (150 mg, 0.63 mmol) andK₂CO₃ (96.6 mg, 0.70 mmol) in acetone (10 mL) was added intermediate11.2 (173 mg, 0.72 mmol). Stirring was continued overnight at roomtemperature. The volatiles were removed under vacuo. The crude was takenup with water, acidified to pH 3 and extracted with EtOAc (3×50 mL). Thetitle compound 23 has been obtained (150 mg, 0.39 mmol) as pure orangesolid after flash chromatography purification eluting with CH₂Cl₂/MeOH(5% for product) and prior titration with hot EtOAc. Yield 62%. ¹H NMR(400 MHz, DMSO) δ 3.92 (s, 2H), 6.72 (t, J=7.7 Hz, 1H), 6.79 (d, J=7.6Hz, 1H), 6.88 (t, J=7.3 Hz, 1H), 7.20 (t, J=4.26 Hz, 1H), 7.77 (d, J=4.7Hz, 1H); 7.92 (d, J=7.8 Hz, 1H), 8.0 (d, J=3.5 Hz, 1H); 9.59 (s, 1H),9.80 (s, 1H); ¹³C NMR (100 MHz, DMSO) δ 35.12, 86.2, 115.5, 119.2,120.1, 121.0, 124.4, 127.0, 128.7, 129.2, 131.4, 141.8, 147.3, 159.2,167.9, 169.1, 170.5. HPLC 97.7%.

II. Biological Activity

Example 29 Determination of ACMSD1 Inhibition

The activity of compounds 1-19 and 21-23 as inhibitors of ACMSD1 wasdetermined by measuring the conversion of 30H-Anthranilic Acid intoproduct (i.e., ACMS) in a spectrophotometrical in vitro assay.

The pre-assay mixture consisting of 3-hydroxyanthranilic acid (3OH-HA),3-hydroxyanthranilic acid, 3,4-diOxygenase (HAO), and a dialyzed crudeextract of E. coli BL21 (DE3) cells expressing the recombinant enzyme,was incubated at 25° C. with monitoring of the increase in absorbance at360 nm due to the formation of ACMS from 3OH-HA. After the reaction wascompleted within ˜2 mins, an aliquot of ACMSD1 solution (prepared andpurified from Pichia Pastoris overexpressing the recombinant enzyme) wasadded, and the decrease in absorbance at 360 nm was followed at 15second intervals. The effect of ACMS concentration on the enzymeactivity was investigated by varying 3OH-HA concentration from 2 to 20μM. Kinetic parameters were calculated from the initial velocity data byusing the Lineweaver-Burk plot.

The rate of the decrease in absorbance caused by ACMSD1 was calculatedby subtracting that of the control reaction mixture without ACMSD fromthat described above. One unit of ACMSD activity was indicated as theamount of enzyme that converts 1 mmol of ACMS per minute at 25° C. Theabsence or a reduction of ACMSD1 activity (e.g., by using ACMSDinhibitors) results in a slow ACMS-spontaneous degradation (i.e.,cyclization to form quinolic acid).

The enzymatic activity was determined at a HAA concentration of 10 μM inthe presence of the compounds in Table 1 below. The compounds weretested at the concentration of about 5 μM and 10 μM and the IC₅₀ wascalculated for compounds showing inhibitory activity higher than 50%.The results are shown in Table 1.

TABLE 1 Compound Activity No. Structure hACMSD IC₅₀ 1

0.050 2

0.066 3

0.031 4

0.012 5

0.049 6

0.077 7

0.082 8

0.082 9

0.96 10

1.7 11

0.071 12

0.088 13

0.136 14

0.74 15

0.76 16

0.11 17

0.005 18

0.010 19

0.025 21

1.99 22

1.1 23

4.9

Example 30 Determination of ACMSD-1 Modulation in HEK293T Cells

HEK293T cells (ATCC) were seeded in six-well plates and transfectedusing Fugene HD to express transiently ACMSD. 24 hrs post transfection,the cells were stimulated for 48 hrs to 72 hrs with differentconcentrations of Compound 1 and then lysed to measure the ACMSDactivity, by measuring the conversion of 30H-Anthranilic Acid intoproduct (i.e. α-amino-beta-carboxymuconate-ε-semialdehyde, ACMS) in aspectrophotometrical in vitro assay. The amount of the whole proteincontent in cell lysates was detected by Bradford analysis. This valuewas used to get the specificity activity of the enzyme normalized in allsamples (mU/ml or ΔE/Δt/mg of total protein).

ACMSD-1 enzyme is known to be expressed in liver, kidney and brain;available cell lines for these cell types were therefore tested todetermine the expression levels of ACMSD. We determined that ACMSD-1 isnot expressed in transformed cell lines from liver and kidney, such asHepG2, HEK293T, Hep3B etc. Transfection of ACMSD was performed toexpress the enzyme in different cellular backgrounds such as COS-7,HEK293T, and HepG2. The HEK293T cellular background proved to be thebest system, with the highest protein production allowing robustmeasurement ACMSD1 enzyme activity. This is probably due to the bettertransfection efficacy observed in HEK293T.

Having determined the optimum stimulation time and transfection protocolcells were stimulated with different concentrations of Compound 1 (about50 nM to about 5 uM). Compound 1 inhibited ACMSD-1 activity, in a dosedependent manner, in this over-expression cell-based assay.

Example 31 Determination of NAD⁺ Content in Human Primary HepatocytesTreated with Compound 4

The NAD⁺ concentration or content was determined in human primaryhepatocytes treated with Compound 4. Vehicle (NT) was used as a control.

At least three experiments were run treating primary hepatocytes withdifferent concentrations of Compound 4 (0.5 μM and 5 μM) after 48 hrsfrom seeding. Compound 4 was replaced every 24 hrs, and then cells weredirectly harvested and lysed with ACN/H₂O (ratio 5:1). LCMS/MS was usedto detect and measure NAD⁺ concentration/content. Screening data showedthat Compound 4 inhibits ACMSD-1 enzyme at concentrations as low as 0.5μM and 5 μM. (FIG. 1)

Example 32 Determination of NAD⁺ Content in Human Primary HepatocytesTreated with Compound 1

The NAD⁺ concentration or content was determined in human primaryhepatocytes treated with Compound 1 and MEHP, a known ACMSD inhibitor.MEHP was used as a control.

At least three experiments were run treating primary hepatocytes withdifferent concentrations of Compound 1 (0.5 μM, 5 μM, and 50 μM) after48 hrs from seeding. Compound 1 was replaced every 24 hrs, and thencells were directly harvested and lysed with ACN/H₂O (ratio 5:1).LCMS/MS was used to detect and measure NAD⁺ concentration/content.Screening data showed that 500 μM of MEHP inhibits 70% of purifiedACMSD-1 enzyme, and that 0.5 μM of Compound 1 has similar inhibitionactivity as 250 μM of MEHP. (FIG. 3)

Example 33 Modulation of SOD2 Activity in AML-12 Cells and MurinePrimary Hepatocytes

The modulation of SOD-2 activity in AML-12 cells and murine primaryhepatocytes treated with either Compound 1 or 17 was measured.

The mouse hepatocytes cell line AML-12 (alpha mouse liver 12) wasobtained from ATCC and grown at 37° C. in a humidified atmosphere of 5%CO₂/95% air in Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12(DMEM/F-12) supplemented with 0.005 mg/ml insulin, 0.005 mg/mltransferrin, 5 ng/ml selenium, 40 ng/ml dexamethasone and 1% gentamycin.ACMSD inhibitor was initially diluted from powder in DMSO to a stockconcentration of 1 mM. This stock was further diluted with water to aconcentration of 100 μM which was used for the cell treatments.

Primary hepatocytes were prepared from 8-12-week-old C57BL/6J mice bycollagenase perfusion method. Mouse livers were perfused with Hank'sbalanced salt solution (HBSS, KCl, 5.4 mM; KH₂PO₄, 0.45 mM; NaCl, 138mM; NaHCO₃, 4.2 mM; Na₂HPO₄, 0.34 mM; glucose, 5.5 mM; HEPES, 1 M; EGTA,50 mM; CaCl₂, 50 mM; pH 7.4). Livers were then washed at a rate of 5ml/min through the portal vein. After washing, livers were perfused withcollagenase (0.025%) solution. Cell viability was assessed by the trypanblue method. Isolated primary hepatocytes were plated with DMEM medium(Gibco) including 10% FCS, 10 units per ml penicillin and HEPES forbuffering. The cells were maintained in culture at 37° C. in ahumidified atmosphere of 5% CO₂/95% air. After 6-8 hrs of attachment,this medium was replaced with media containing different concentrationsof an ACMSD inhibitor (i.e., Compound 1 or Compound 17) or with thecorresponding concentration of DMSO (as a control). Primary hepatocyteswere harvested about 24 hrs later if not indicated differently.

Primary hepatocytes or AML-12 cells were then lysed in a 20 mM HEPESbuffer (Gibco), pH 7.2, containing 1 mM EGTA (Sigma), 210 mM mannitol(Sigma), and 70 mM sucrose (AMRESCO). Total protein concentration wasdetermined using the Bradford assay (BioRad). SOD-2 activity wasdetermined at indicated times after ACMSD inhibitor treatment by the SODAssay Kit (Cayman Chemical) according to the manufacturer'sinstructions. In order to specifically detect the SOD2 activity 2 mMpotassium cyanide was added to the assay, which inhibited both Cu/Zn-SODand extracellular SOD, resulting in the detection of only Mn-SOD (SOD-2)activity. Absorbance was determined with a Victor X4 multi-label platereader (Perkin-Elmer) at 450 nm. Results are expressed in U/ml/mg ofprotein according to the standard curve and measured proteinconcentration.

The oxidative stress resistance pathway, which seemed to be induced uponACSMD inhibition, was explored by measuring the activity of SOD2. Theresults showed that SOD2 was induced in a dose-dependent manner in bothAML-12 and primary murine hepatocytes by both Compound 17 andCompound 1. In primary hepatocytes, which express ACMSD at significantlyhigher levels than AML-12, effects were observed at a dose of about 5 nMand reached a maximum at dose of about 50 nM. Both Compound 17 andCompound 1 were able to induce the activity of SOD2 in a dose-dependentmanner (FIG. 5A and FIG. 5B)

Example 34 Determination of NAD⁺ Content in Murine Primary Hepatocytes

NAD⁺ levels were determined in human primary hepatocytes treated withCompound 17.

NAD⁺ was extracted using acidic extraction method. Samples werecollected and homogenized in 70% ice-cold perchloric acid (HClO₄). Afterinsoluble protein parts were pelleted by adding potassium carbonate(K₂CO₃), the samples were separated by high-performance liquidchromatography (HPLC) and analyzed by mass-spectrometry. The proteins inthe pellet were quantified by Bradford assay and were used fornormalization.

The exposure of primary hepatocytes to 5 nM, 10 nM and 50 nM of theACMSD inhibitor Compound 17 for 24 hours induced a significant anddose-dependent increase in intra-cellular NAD⁺ levels. A significanteffect on NAD⁺ levels was observed at concentrations as low as 5 nMconcentration. (FIG. 2)

Example 35 RT-qPCR Analysis of SIRT1-Regulated Genes in AML-12 Cells,Hepa-1.6 Cells and Primary Murine Hepatocytes Treated with Compound 1 or17

Gene expression of ACMSD and genes known to be regulated by SIRT1, (anenzyme that is strictly NAD⁺ dependent) such as Pgc1a, Sod1, Sod2(MnSOD), were analysed in AML-12 cells, Hepa-1.6 cells and primarymurine hepatocytes treated with Compound 1 or 17.

Cells (AML-12, Hepa-1.6, HEK-293, primary human and murine hepatocytes)were treated with different concentrations of Compound 1 or Compound 17.Total RNA was extracted from cells using TRIzol (Invitrogen) accordingto the manufacturer's instructions. The RNA was treated with DNase, and2 μg of RNA was used for reverse transcription (RT). 50× diluted cDNAwas used for RT-quantitative PCR (RT-qPCR) reactions. The RT-qPCRreactions were performed using the Light-Cycler system (Roche AppliedScience) and a qPCR Supermix (QIAGEN) with the indicated primers. Theaverage of at least three technical repeats was used for each biologicaldata point.

A dose-dependent increase in mRNA expression levels of genes known to beregulated by SIRT1, (an enzyme that is strictly NAD⁺ dependent) such asPgc1a, Sod2 (MnSOD), but not Sod1 (Cu—Zn SOD), was observed in primarymouse hepatocytes treated for 24 hrs with Compound 17 (5 nM-500 nMrange). The observed increase in the gene expression was dose-dependent,which is in line with the dose-dependent increase in SOD2 enzymaticactivity observed in Example 32 (FIG. 5). Sod2 mRNA levels were alsoincreased in a dose-dependent manner in the AML-12 cells and Hepa-1.6hepatic cell lines after 24 hrs of treatment with Compound 1. Thesechanges in mRNA expression are compatible with the activation of SIRT1,subsequent to the induction in NAD⁺ levels by inhibition of ACMSD1activity by Compound 17. (FIG. 4 and FIG. 5)

Example 36 Modulation of Caspase 3/7 Activity in MDCK Cells

An in vitro study was performed to determine the effects of compounds ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof, on Acute Kidney Injury inMDCK cells.

MDCK cells (MDCK (NBL-2) ATCC® CCL-34™) were cultured in base mediumATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003with fetal bovine serum (FBS) to a final concentration of 10%. 10,000cells were plated into 96 wells and 24 hours after cell plating themedium was changed with fresh medium supplemented with 1% FBS. Cisplatin(50 μM for 16 hrs) was then used to induce cell injury. Differentconcentrations of Compound 18 (in 1% DMSO) were added in combinationwith cisplatin (FIG. 1) or 1 hour prior adding cisplatin (FIG. 2).

Caspase 3/7 activity (Promega) was determined according to standardprocedures using a luminescent signal readout on a Victor V plate reader(PerkinElmer). Each experiment/condition was performed in triplicate.

Caspase activity was analyzed as percentage effect normalized to thecisplatin alone (100%) and vehicle treated cells as 0% of caspaseactivity. Data were analyzed by GraphPad Software. One-way analysis ofvariance (Dunnett's Multiple Comparison test) was used for statisticalanalyses.

As shown in FIG. 1, MDCK cells were treated with differentconcentrations of Compound 18 (0.01 μM to 100 μM) in combination withcisplatin (cisp). The EC₅₀ value was calculated as equal to 70 μM. Cellstreated with Compound 18 decreased caspase activity in significantmanner at a concentration of 100 μM compared to cells treated withcisplatin alone (p<0.001).

MDCK cells were also treated with different concentrations (1 μM to 125μM) of Compound 18 one hour prior to the addition of cisplatin (cisp).As shown in FIG. 2, cells treated with Compound 18 decreased caspaseactivity in significant manner at a concentration of about 30 μM toabout 125 μM compared to cells treated with cisplatin alone (p<0.001).The EC₅₀ value was calculated as equal to 30 μM.

Data show that Compound 18 decreases, in significant manner, theactivity of caspase 3/7 induced by cisplatin (FIG. 1) the protectiveeffect is particularly noteworthy if Compound 18 is added before insultwith the injury agent (cisplatin) as shown in FIG. 2.

Example 37 Cytotoxicity and hERG Screening

Cytotoxicity: 20000 HePG2 and AML-12 cells were seeded in 96 well plate(Viewplate PerkinElmer). Dose-response of the compound in Table 2 wasperformed using HP D300 digital dispenser, ranging from 10 nM to 300 μMwith constant DMSO 1% in medium. Cells were stimulated for 4 hrs at 37°C.; the supernatant was used to perform LDH release (Cytotox-one,Promega) as a measure of necrosis while the cells were lysed to detectATP level for determining cell viability (Celltiter-glo, Promega)according to manufacturer's instructions.

The Predictor hERG assay kit (Invitrogen), containing membranepreparations from Chinese hamster ovary cells stably transfected withhERG potassium channel and a high-affinity red fluorescent hERG channelligand (tracer), was used for the determination of hERG channel affinitybinding of the test compounds in Table 2. Compounds that bind to thehERG channel protein (competitors) were identified by their ability todisplace the tracer, resulting in a lower fluorescence polarization. Thefinal concentration of DMSO in each well was maintained at 1%. Theassays were performed according to the manufacturer's protocol(Invitrogen).

The results are shown in Table 2.

TABLE 2 Cytotoxicity Compound Compound Structure AML-12 Hep-G2 hERG 17

Not Toxic Not Toxic No Activity 8

Not Toxic Not Toxic No Activity 4

Not Toxic Not Toxic No Activity 19

Not Toxic Not Toxic No Activity 5

Not Toxic Not Toxic No Activity 1

Not Toxic Not Toxic No Activity 11

Not Toxic Not Toxic No Activity

Example 38 C. elegans Experiments—ACMSD1 Silencing, Lifespan Assays,Mobility Assessment and GFP Fluorescence Quantification

C. elegans (Caenorhabditis elegans) strains were provided by theCaenorhabditis Genetics Center (University of Minnesota). Worms weremaintained on Nematode Growth Medium (NGM) agar plates seeded with E.coli OP50 bacteria at 20° C., unless stated otherwise. The strains usedfor the experiments were the following: Bristol N2, NL2099(rrf-3(pk1426)II), KN259 (huIs33 [sod-3::GFP+pRF4(rol-6(su1006))]).

Bacterial feeding RNAi experiments were carried out as follows: wormswere grown on NGM agar plates containing Carbenicillin and IPTG at finalconcentrations of 25 μg/ml and 1 mM respectively and seeded withbacterial cultures taken from Ahringer library. Clones used were acmsd-1(Y71D11A.3), sir-2.1 (R11A8.4), and daf-16 (R13H8.1). Clones werepurchased from GeneService and their identity was confirmed bysequencing. For the double RNAi experiments bacterial cultures weremixed before seeding on NGM plates. The control RNAi in this kind ofexperiments was 50% diluted with control empty vector RNAi bacteria.

The nematode Caenorhabditis elegans was used as a model system toconfirm the activation of the oxidative stress defence that we haveobserved in cells at the level of an intact organism. The effects ofacmsd-1 RNAi were assessed in C. elegans by RT-qPCR. The total RNA wasextracted from cells using TRIzol (Invitrogen) according to themanufacturer's instructions. RNA was treated with DNase, and 2 μg of RNAwas used for reverse transcription (RT). 50× diluted cDNA was used forRT-quantitative PCR (RT-qPCR) reactions. The RT-qPCR reactions wereperformed using the Light-Cycler system (Roche Applied Science) and aqPCR Supermix (QIAGEN) with the indicated primers. The average of atleast three technical repeats was used for each biological data point.

C. elegans lifespan assays were carried at 20° C. as follows. Animalswere exposed to NAC (N-acetyl cysteine) at a final concentration of 5 mMfrom a 0.5 M aqueous stock from the young adult stage. Sodium pyruvatewas added at a final concentration of 2.5 mM to NGM plates containingcarbenicillin (100 lg mL) and seeded with UV-killed OP50. After 5 daysof RNAi treatment, worms were transferred to plates containing paraquatand seeded with acmsd-1 RNAi bacteria. Control animals were grown duringthe first 5 days of adulthood on RNAi bacteria containing the emptyvector and then transferred to plates containing paraquat and seededwith acmsd-1 RNAi bacteria. Survival analyses were performed using theKaplan-Meier method, and the significance of differences betweensurvival curves was calculated using the log rank test. The statisticalsoftware used was XLSTAT 2007 (XLSTAT, Brooklyn, N.Y., USA), and allP-values <0.05 were considered significant. 100 worms were used percondition and scored every 2 days. The reasons for censoring were the<<exploded vulva>> phenotype or worms that crawled off the plate. Whereindicated, paraquat was added on top of the agar plates at the indicatedconcentration. Once the paraquat solution was completely dried, L4 wormswere transferred to these agar plates and monitored for 5-6 days everyday. By day 6 all the paraquat tests were stopped because a smallpercentage in worm population could start to die naturally and ratherthan dying due to the paraquat effects.

The movement of worms was recorded for 45 seconds at days 1, 3, and 5 ofadulthood using a Nikon DS-L2/DS-Fil camera and controller setup,attached to both a computer and a standard bright field microscope. Foreach condition five plates of worms, with 10 worms per plate were used.The movement of worms during aging was calculated by taking an integralof the speed value which was assessed by following the worm centroidswith a modified version of the freely-available for the Parallel WormTracker for MATLAB.

Fluorescence intensity in worm strains expressing GFP-reporter proteinswas quantified using Victor X4 plate reader (Perkin Elmer). The animalswere prepared in the following way: eighty worms per condition (at thecorresponding ages) were picked (20 worms per well of a black-walled96-well plate) and placed into the M9 medium. Each experiment wasrepeated at least twice.

The expression level of acmsd-1 mRNA was significantly reducedconfirming the efficacy of RNAi mediated gene knock-down (FIG. 7A). Theworm ortholog of MnSOD, SOD-3, was induced at its mRNA level withconcomitant downregulation of the acmsd-1 gene with RNAi. A significantincrease at the protein level of SOD-3 was also observed at Day 3 ofadulthood. (FIG. 7B) ACMSD downregulation improved worm lifespan andthis improvement was SIR-2.1- and DAF-16-dependent. (FIG. 7C)

Moreover, worms exposed to acmsd-1 RNAi lived longer and showed improvedperformance in mobility assays when treated with paraquat, a well-knownROS inducer that is widely used to mimic oxidative stress in C. elegans.(FIGS. 7D and 7E)

The better survival at paraquat conditions was independent on thedevelopmental stage at which worms were exposed to acmsd-1 RNAi. (FIG.7F)

This increase in lifespan under oxidative stress conditions was nolonger observed when DAF-16, the worm ortholog of FoxO1, wasdownregulated, meaning that better oxidative stress resistance wasDAF-16 dependent. (FIG. 7G)

Example 39 Study of the Anti-Diabetic Effects of Compounds of Formula(I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III) inC57BL/6J and KK-Ay Mice

A glucose tolerance test is performed on male C57BL/6J and KK-Ay mice todetermine the effects of compounds of Formula (I), Formula (Ia), Formula(Ib), Formula (II), or Formula (III) on glucose and insulin levels.

Male C57BL/6J and KK-Ay mice, 6-7 weeks of age, are obtained, e.g., fromCharles River Laboratories France and CLEA Japan, respectively. Mice arefed from the age of 8 weeks onwards with regular chow (CD-Harlan 2018),a high fat diet (HFD-Harlan 06414). A compound of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, is mixed with the HFD at 180mg kg⁻¹ of food. On the basis of their daily food intake, this resultsin a daily dose of about 15 mg kg⁻¹ body weight. The mice are fasted for4 hrs before blood and tissues are harvested for RNA isolation, lipidmeasurements and histology. Oxygen consumption is measured with theOxymax apparatus (Columbus Instruments). Histological analysis andtransmission electron microscopy are performed.

An oral glucose tolerance test is performed in the animals that arefasted overnight. Glucose is administered by gavage at a dose of 2 g/kg.An intraperitoneal insulin tolerance test is performed in animals fastedfor 4 hrs. Insulin is injected at a dose of 0.75 U/kg body weight.Glucose is quantified with the Maxi Kit Glucometer 4 (Bayer Diagnostic)or Glucose RTU (bioMerieux Inc.) and plasma insulin concentrations aremeasured by ELISA (Cristal Chem Inc.). Statistical differences aredetermined by either ANOVA or Student's t-test.

Example 40 Study of the Anti-Diabetic and Obesity Effects of Compoundsof Formula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula(III) in Db/Db Mice with LepR Mutation

A study of the anti-diabetic effects of the compounds of Formula (I),Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, is conducted in geneticallyobese Leprdb/J (db/db) mice.

Animals are bred and housed in a temperature- and humidity-controlledenvironment in compliance with FELASA-protocols. From an age of threeweeks, mice are fed a high-fat diet (HFD) (Harlan 06414). Mostpharmacological studies are started in diabetic eight-week-old db/db andwild type (wt) references.

Subchronic Intervention

db/db mice are treated once/day with a compound of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, for 14 days between 5-6 PMbefore dark-phase onset (6 PM). Blood samples are collected after 4 hrsof fasting the mice prior to the first dose and at 18±2 hrs after thelast dose. Glucose concentrations of each blood sample are determined.

Acute Intervention Glucose

Initial blood samples are collected in random-fed db/db mice between 6-8AM after light-phase-onset (6 AM), then compounds of Formula (I),Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, are administered, diet-accessis restricted, and the second blood sample is collected 4 hrspost-treatment. Thereafter, mice are subjected to an oral glucosetolerance test (OGTT1: 1 g glucose/kg body mass) and blood glucoseconcentrations are determined at 0.5, 1, 2, 3, and 4 hrs after eachglucose challenge.

Euglycemic-hyperinsulinemic Clamps Assay

db/db mice receive a permanent jugular vein catheter underketamine/xylazine anesthesia. For six to seven days, later (after 6 AM)food-access is restricted. Conscious mice are placed in oversizedrat-restrainers and warmed by warming pads. Catheter-ends are thenconnected to syringes in CMA402-pumps (Axel Semrau, Sprockhoevel,Germany). After 110 minutes of primed-continuous [3-³H]glucose infusion(1.85 kBq/min), a blood sample is collected to determine plasma insulin,glucose and [3-³H]glucose concentrations and to calculate basalendogenous glucose appearance rates. The mice then receive vehicle or acompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof, viagavage.

Subsequently, glucose-1 clamps are started with a [3-³H]glucose infusion(3.7 kBq/min) containing insulin (36 pmol/kg*min⁻¹; HumulinR, Lilly,USA) causing a moderate net-increase in plasma insulin concentrations.Blood glucose concentrations are measured every 10 minutes and targetglycemia is established by adjusting the rate of a 20% glucose infusion(GIR). At minute 120, 2-deoxy-D-[1-¹⁴C]glucose (370 kBq) is givenintravenously. Blood samples are collected at minute 30, 60, 90, 100,110, 120, 122, 125, 130, and 140. The mice are then sacrificed (i.e.,through an intravenous ketamine/xylazine-overdose). Gastrocnemius muscleand epididymal adipose tissue are collected, immediately snap-frozen inliquid nitrogen, and stored at −80° C. 2-[¹⁴C]deoxyglucose-6-phosphateis extracted from the tissue and glucose uptake rates (Rg) arecalculated.

Plasma [³H]- and [¹⁴C]-radioactivity is determined in deproteinizedplasma after [³H₂O] evaporation. Glucose fluxes under basal conditionsand between glucose clamp minute 60 to 90 and 90 to 120 are estimated asfollows: whole-body glucose disappearance rate (Rd)=[3-³H]GIR(dpm/min)/plasma [3-³H]glucose specific activity (dpm/min*mol); basalEndo Ra=[3-³H]GIR (dpm/min)/plasma [3-³H]glucose specific activity(dpm/min*mol); glucose-clamp Endo Ra=GIR-Rd. Ultima-Goldscintillation-cocktail, radioisotopes, and a Tri-Carb2910TR are obtainedfrom Perkin Elmer (Germany).

Assays from Blood, Plasma, Urine

Blood samples are collected from lateral tail veins. Blood glucose ismeasured with a glucometer (Contour, Bayer Vital, Germany), urine andplasma glucose with a colorimetric Glucose LabAssay (Wako, Germany), andHbA1c with A1cNow+ (Bayer Vital) or Clover Analyzer (Inopia, SouthKorea).

Analyses of Disease Onset and Survival

Disease onset is defined as the last day of individual peak body weightbefore gradual loss occurs. The stages of disease are defined asfollows: the early stage of disease is defined as the duration of timebetween peak body weight until loss of 10% of peak body weight. The latestage of disease is defined as the duration of time between 10% loss ofpeak body weight until the end stage of disease. The end stage ofdisease is defined as the day when an animal could no longer rightitself within 30 s for three consecutive trials when placed on its side.Animals are euthanized at the end stage of disease.

Body Composition Measurements

Body weights are assessed weekly for at least 13 weeks. Brown adiposetissue (BAT) and gonadal white adipose tissue (WAT) are dissected andweighed at the indicated age. Total lean mass, % of WAT and BMD (bonemineral density) are determined by DEXA (PIXImus DEXA; GE).

Indirect Calorimetry, Food Intake and Activity

Animals are initially weighed and acclimated to the test cage. Volumeoxygen (VO₂) and volume carbon dioxide production (VCO₂) are measuredevery 20 min using the Oxymax Comprehensive Laboratory Animal MonitoringSystem (CLAMS) (Columbus Instruments) and are reported as average VO₂per hour normalized to body weight (mL/h/kg). Using the CLAMS machine,activity counts by infrared beam interruptions and food intake aresimultaneously measured. More specifically, food intake is measured bydeducting the weight of powderized food pellets at the end ofexperimentation from the starting weight at the beginning ofexperimentation. To complement this experiment and to control for anovel environment that may affect feeding behaviour, we also perform amore ‘manual’ experiment, wherein a set weight of food pellets is placedat the same time each day into a clean home cage, which holds a mouse.The next day the weight of the remaining pellets is recorded anddeducted from the starting weight. This experiment is performed for 14days straight. The body weight of each mouse is also recorded daily.Results for each genotype are similar to that acquired from the CLAMS.

Statistical Analyses.

Considering a 1-β larger than 0.9 statistically powerful, we estimateappropriate group numbers from pilot studies a priori. One- or two-wayAnalyses of Variance (Bonferroni post-tests) or t-tests are performed.

Example 41 Study of the Effects of Compounds of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III) on Non-AlcoholicFatty Liver Disease (NAFLD) and Non-Alcoholic Steatohepatitis (NASH) inMice

A study is performed to determine the effects of compounds of Formula(I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, on non-alcoholic fatty liverdisease (NAFLD) and non-alcoholic steatohepatitis (NASH) in maleC57BL/6J fed a high fat and high sucrose diet.

Male C57BL/6J mice (The Jackson Laboratory, Bar Harbor, Me., USA) arehoused under a 14 hrs light-10 hrs dark cycle at 21-23° C. and have adlibitum access to water during the entire experiment. From the age of 6weeks, mice are fed a ‘Western’ HF-HSD with 44.6% of kcal derived fromfat (of which 61% saturated fatty acids) and 40.6% of kcal derived fromcarbohydrates (primarily sucrose 340 g/kg diet) (TD.08811, 45% kcal FatDiet, Harlan Laboratories Inc., Madison, Wis., USA) or normal chow diet(NCD) as control (V1534-000 ssniff R/M-H, ssniff Spezialdiaten GmbH,Soest, Germany). The animals are then treated with a compound of Formula(I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, or a control for 4, 12 or 20weeks (n=8 per group for every time point), after which they aresacrificed.

Body weight and food intake are monitored weekly on the same day. Aftersedation with sodium pentobarbital (intraperitoneal injection, 50 mg/kgbody weight), total fat mass is analysed by dual-energy X-rayabsorptiometry (DEXA) (PIXImus densitometer, Lunar Corp., Madison, Wis.,USA). Intraperitoneal glucose tolerance test (IPGTT) is performed in 6hrs fasted mice. Tail vein glucose levels are measured with a BayerContour glucometer immediately before (time point 0 min) and 15, 30, 60,90 and 150 min after glucose administration (1 g glucose/kg bodyweight). Insulin resistance is calculated using the Homeostasis Model ofInsulin Resistance (HOMA-IR) index: (fasting insulin (ng/mL)×fastingglucose (mg/dL))/405.

Sacrifice

After a 6 hrs fasting period, mice are anaesthetised with sodiumpentobarbital (intraperitoneal injection, 50 mg/kg body weight) andsacrificed by blood sampling via cardiac puncture. Plasma is obtained bycentrifugation of blood (6000 rpm for 5 min at 4° C.) that is collectedin heparinised syringes. Tissues are either snap frozen in liquidnitrogen or stored at −80° C. together with the plasma until furtherbiochemical and molecular analyses or preserved for histologicalanalysis.

Histological Analyses

Liver samples are routinely fixed in buffered formalin (4%) and embeddedin paraffin. Serial 4 mm thick sections are stained with H&E andpicrosirius red to assess fibrosis. Frozen liver sections are stainedwith Oil Red O to assess lipid accumulation. All liver biopsies areanalysed by an expert liver pathologist, blinded to the dietarycondition or surgical intervention. Steatosis, activity and fibrosis aresemiquantitatively scored according to the NASH-Clinical ResearchNetwork criteria. The amount of steatosis (percentage of hepatocytescontaining fat droplets) is scored as 0 (<5%), 1 (5-33%), 2 (>33-66%)and 3 (>66%). Hepatocyte ballooning is classified as 0 (none), 1 (few)or 2 (many cells/prominent ballooning). Foci of lobular inflammation arescored as 0 (no foci), 1 (<2 foci per 200x field), 2 (2-4 foci per 200xfield) and 3 (>4 foci per 200x field). Fibrosis is scored as stage F0(no fibrosis), stage F1a (mild, zone 3, perisinusoidal fibrosis), stageF1b (moderate, zone 3, perisinusoidal fibrosis), stage F1c(portal/periportal fibrosis), stage F2 (perisinusoidal andportal/periportal fibrosis), stage F3 (bridging fibrosis) and stage F4(cirrhosis). Diagnosis of NASH is based on accepted histologicalcriteria. Severity of the disease is assessed using the NAS (NAFLDactivity score) as the unweighted sum of scores of steatosis, hepatocyteballooning and lobular inflammation. Percentage of fibrosis isquantitated by morphometry from digitalised sirius red stained sectionsusing the Aperio system after tuning the threshold of fibrosis detectionunder visual control. Results are expressed as collagen proportionalarea.

Example 42 Study of the Effects of Compounds of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III) on Non-alcoholicFatty Liver Disease (NAFLD) and Non-alcoholic Steatohepatitis (NASH) inMethionine and Choline Deficient Mice

A study is performed to determine the effects of compounds of Formula(I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, on non-alcoholic fatty liverdisease (NAFLD) and non-alcoholic steatohepatitis (NASH) in malewildtype mice fed a methionine- and choline-deficient diet.

Wildtype mice housed in 12-hour light/dark cycles, with free access tofood and water are used. At least 5 animals per time point are analysed.All experiments are repeated at least three times. For dietarytreatment, 8-12 weeks old male mice weighing 25 g are either fed amethionine- and choline-deficient diet (MCD to induce NASH) or chow diet(as a control). Animal experiments and evaluation of NAFLD and NASH asdescribed above in Example 40 for mice fed the high fat and high sucrosediet.

Example 43 Study of the Effects of Compounds of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III) on Atherosclerosis inHigh Cholesterol Fed LDL-R Knockout Mice

A study is performed to determine the effects of compounds of Formula(I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, on atherosclerosis in highcholesterol fed LDL-R knockout mice.

LDL-R knockout (KO) mice are backcrossed for ten generations with theC57BL/6J strain, yielding congenic C57BL/6J animals. The controls thatare used are littermates in all experiments. The animals are treatedwith a compound of Formula (I), Formula (Ia), Formula (Ib), Formula(II), or Formula (III), or pharmaceutically acceptable salt thereof, ora control. Mice are sacrificed 12 weeks after the initiation of theatherogenic diet (TD94059; Harlan), after which the heart and aorta areperfused with PBS and subsequently fixed (Shandon Formal Fixx, ThermoScientific). Atherosclerosis is assessed by an Oil red O staining of theaortic root and quantified with MetaMorph software. Biochemistryparameters are measured with the appropriate kits in the COBAS C111(Roche). For the in vivo lipopolysaccharide (LPS) study, mice areintraperitoneally injected with 100 mg of LPS, and blood is taken fromthe tail vein. TNFα levels are quantified with Mouse TNFα ELISAReady-SET-Go! (eBioscience) assay. Blood cell counts are determined withAdvia2120 (Siemens Healthcare Diagnostics).

The Student's t test is used to calculate the statistical significance.In case of multiple testing (i.e., the comparison of more than twogroups), this test is preceded by the ANOVA test. P<0.05 is consideredstatistically significant. Results represent the mean±SEM.

Example 44 Study of the Effects of Compounds of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III) on InheritedMitochondrial Disease in Sco2^(KO/KI) Mice

A study is performed to determine the effects of compounds of Formula(I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, on inherited mitochondrialdisease in Sco2^(KO/KI) mice.

Anti-COI, anti-COX5a, anti-Ndufa9, anti-SDH-HA, and anti-Core 2 are fromInvitrogen; anti-GAPDH is from Millipore; anti-FoxO1 andanti-acetylated-FoxO1 are from Cell Signaling and Santa Cruz,respectively. Anti-mouse secondary antibodies are from Amersham.Chemicals are from Sigma. Oligonucleotides are from PRIMM, Italy.

Compounds of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof, aredissolved in water and added to a standard powder diet (Mucedola, Italy)at the appropriate concentration of 50 mg/Kg/day. Pellets containing thecompounds of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof, or thevehicles are reconstituted by hand and kept frozen at −20° C. untilneeded. The diet supply is changed every three days, and only the amountneeded is thawed at each time and administered ad libitum for one month.Sco2^(KO/KI) mice are maintained in a temperature- andhumidity-controlled animal-care facility, with a 12 hrs light/dark cycleand free access to water and food. Animals are sacrificed by cervicaldislocation.

Morphological Analysis

For histochemical analysis, tissues are frozen in liquid-nitrogenprecooled isopentane. Series of 8 mm thick sections are stained for COXand SDH.

Biochemical Analysis of MRC Complexes

Muscle quadriceps samples stored in liquid nitrogen are homogenized in10 mM phosphate buffer (pH 7.4), and the spectrophotometric activity ofcI, cII, cIII, and cIV, as well as CS, is measured as described. Notethat in all panels the activity of cII is multiplied by 10 forvisualization clarity.

NAD⁺ Determination

NAD⁺ is extracted using acidic and alkaline extraction methods,respectively. Tissue NAD⁺ is analysed with mass spectrometry aspreviously described.

Example 45 Study of the Effects of Compounds of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III) on InheritedMitochondrial Disease in Deletor Mice

A study is performed to determine the effects of compounds of Formula(I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, on inherited mitochondrialdisease in Deletor mice.

The Deletor mouse model is generated in C57BL/6 congenic background andhas been previously characterized (Tyynismaa et al, 2005); WT mice arelittermates from the same congenic mouse strain C57BL/6J. Deletor and WTmale mice are administered either chow diet (CD) or a compound ofFormula (I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III),or a pharmaceutically acceptable salt thereof, admixed with the CD atthe appropriate concentration. The food pellets are manually prepared bymixing a compound of Formula (I), Formula (Ia), Formula (Ib), Formula(II), or Formula (III), or a pharmaceutically acceptable salt thereof,into the powdered food as described for the Sco2^(KO/KI) mice in Example43 and stored at −20° C. The mice are housed in standard animalfacility, under a 12 hrs dark/light cycle. They have ad libitum accessto food and water. The pre-manifestation group consists of 12 Deletorsand 12 WT mice, and the post-manifestation group of 24 Deletors and 24WT mice, receiving either a compound of Formula (I), Formula (Ia),Formula (Ib), Formula (II), or Formula (III), or a pharmaceuticallyacceptable salt thereof, or CD diet. During the intervention, the miceare regularly monitored for weight, food consumption, and physicalendurance. Their exercise capability is measured twice by treadmillexercise test (Exer-6M Treadmill, Columbus Instrument) at the start andthe end of the diet. The exercise test protocol consists of the initialrunning speed of 7 m/s which is increased every 2 min by 2 m/s andcontinued until the animal is unable to run or repeatedly falls from thebelt at the stimulus site.

Oxygen consumption and carbon dioxide production, as well as spontaneousmoving and feeding activities, are recorded by Oxymax Lab AnimalMonitoring System (CLAMS; Columbus Instruments, OH, USA). The mice arekept in individual cages inside a CLAMS chamber for 3 days; the firstday and night is a nonrecording adjustment period followed by a 24 hrsrecording at thermoneutrality (+30° C.). The results of O₂ consumptionand CO₂ production are used to calculate respiratory exchange rate andanalysed separately from the light (inactive) and dark (active) periodsof the day.

Morphologic Analysis

Tissue sections are prepared from the quadriceps, liver, and BAT.Samples are embedded with OCT Compound Embedding Medium (Tissue-Tek) andsnap-frozen in 2-methylbutane in liquid nitrogen. Frozen sections (12lm) from quadriceps are assayed for in situ histochemical COX andsuccinate dehydrogenase (SDH) activities simultaneously. The activitiesfrom the quadriceps sections, the COX-negative and the COX-negative plusSDH positive and normal fibres are calculated. Approximately 2000 fibresare counted from each mouse sample. The intensity of COX histochemicalactivity from quadriceps for both oxidative and non-oxidative fibres ismeasured with Image J software. Frozen sections (8 μm) from liver andBAT are stained with Oil Red O. For plastic embedding, quadriceps,liver, and BAT samples are fixed in 2.5% glutaraldehyde, treated with 1%osmium tetroxide, dehydrated in ethanol, and embedded in epoxy resin.Semi-thin (1 μm) sections are stained with methyl blue (0.5% w/v) andboric acid (1% w/v). The interesting areas for the ultrastructuralanalyses are selected by inspection of the light microscopic sections.For transmission electron microscopy, ultrathin (60-90 nm) sections arecut on grids and stained with uranyl acetate and lead citrate and viewedwith a Transmission Electron Microscope. Crista content in both BAT andmuscle is determined from electron micrographs, utilizing a 1 μm“intra-mitochondrial measuring stick,” placed perpendicular to cristae.Skeletal muscle samples are also analysed for citrate synthase activity.

Example 46 Study of the Effects of Compounds of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III) on Kidney Disease

A study is performed to determine the effects of compounds of Formula(I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, on kidney disease in C57BL/6JWT mice. (Wei, Q., et al. “Mouse model of ischemic acute kidney injury:technical notes and tricks” American Journal of Physiology-RenalPhysiology, 303(11), F1487-F1494)

C57BL/6J WT mice are purchased from Charles-River. All mice are fed astandard commercial diet while housed at an ambient temperature of20-22° C. with a relative humidity of 50±5% under 12/12 hrs light-darkcycle in a specific pathogen-free facility. The experimental mice are 8weeks old and are divided into four groups: control (n=5); cisplatin (20mg/kg; Sigma Chemical, St Louis, Mo.; n=5); a compound of Formula (I),Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, and cisplatin (n=5); and acompound of Formula (I), Formula (Ia), Formula (Ib), Formula (II), orFormula (III), or a pharmaceutically acceptable salt thereof, alone (40mg/kg; n=5). The dose and time of cisplatin treatment for nephrotoxicityare chosen according to a published method. A compound of Formula (I),Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, is administered orally once aday for 4 days. Cisplatin is injected once at 12 hrs after the firstadministration of a compound of Formula (I), Formula (Ia), Formula (Ib),Formula (II), or Formula (III), or a pharmaceutically acceptable saltthereof. The mice are sacrificed at 72 hrs after the single cisplatininjection.

Assays for Renal Functional Markers and Proinflammatory Cytokines

For renal function analysis, serum is isolated and stored at −80° C.until use. Serum creatinine and BUN levels are measured using an assaykit according to the manufacturer's instructions (BioVision, Milpitas,Calif.). In addition, the proinflammatory cytokines TNF-α, IL-1b, andIL-6 from serum or homogenates from kidney tissue are quantified byELISA (Quantikine Kit; R&D Systems, Minneapolis, Minn.) according to themanufacturer's instructions. For measuring cytokines, kidney tissue ishomogenized in phosphate buffered saline containing 0.05% Tween-20.Aliquots containing 300 mg of total protein are used. A metabolic cageis used for collecting urine to analyse the level of urinary cytokines.The sample size for each group is five.

Alternative Study of the Effects of Compounds of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III) on Kidney Disease

Alternatively, C57BL/6J WT mice are numbered and kept in acclimatizationfor a period of 5-7 days before initiation of the experiment. (Wei, Q.,et al. “Mouse model of ischemic acute kidney injury: technical notes andtricks” American Journal of Physiology—Renal Physiology, 303(11),F1487-F1494) Mice are randomized into different treatment groups basedon their body weight. Different groups are maintained on Harlan diet2916. Mice are then maintained on the respective diets for 10 days priorto bilateral Ischemic kidney injury. Body weight measurement is madeonce at randomization and once on day 7. Food consumption is evaluatedonce on day 7. Blood is collected by retro-orbital puncture under mildIsoflurane anesthesia and used for analysis of basal blood urea nitrogenlevels (BUN) on day 9.

Mice are anesthetized with ketamine (80 mg/kg i.p) and/or Xylazine (10mg/kg, i.p.) and placed on a surgical platform in a dorsal position.Both kidneys are exposed through flank incisions and renal pedicles areoccluded using vascular clamps for 25 minutes. The clamp is then removedand the surgical site is sutured. 1 ml of physiological saline isadministered intra-peritoneally after closing the wound to preventdehydration. The sham-operated group is subjected to similar surgicalprocedures, except that the occluding clamp is not applied. Animals aremonitored until recovery from anesthesia and returned to their homecage. Animals are observed every day for general clinical signs andsymptoms and mortality.

One day prior to termination, animals are individually housed inmetabolic cages for 12 h and urine is collected for estimation of urea,creatinine, sodium and potassium.

On days 12, 14, & 16 blood is collected by retro orbital puncture undermild isoflurane anesthesia and plasma is used for analysis of blood ureanitrogen levels (BUN) and serum creatinine. Animals are then euthanizedby CO₂ inhalation and organs are collected. One kidney is fixed in 10%neutral buffered formalin and the other is flash frozen in liquidnitrogen, stored at −80° C. and used for the estimation of lipidperoxidation, GSH, MPO and SOD levels.

Histological Analysis and Neutrophil Counting

Mouse kidneys are fixed in 4% formaldehyde and embedded in paraffin wax.The 5-mm-thick sections are deparaffinised in xylene and rehydratedthrough graded concentrations of ethanol. H&E and PAS staining areperformed using standard protocols. Images are collected and analysedusing a light microscope (IX71, Olympus, Tokyo, Japan) with DP analysersoftware (DP70-BSW, Tokyo, Japan). Tubular damage in PAS-stained kidneysections is examined under a light microscope and scored based on thepercentage of cortical tubular necrosis: 0=normal, 1=1-10, 2=11-25,3=26-45, 4=46-75, and 5=76-100%. Slides are scored in a blinded manner,and results are means±s.d. of 10 representative fields/group. Severitycriterion for tubular necrosis displaying the loss of the proximaltubular brush border and cast formation are used to classify samples.The sample size for each group is 10. Neutrophil infiltration isquantitatively assessed on PAS stained tissue by a renal pathologist bycounting the number of neutrophils per high-power field (×400). At least10 fields are counted in the outer stripe of the outer medulla for eachslide.

All values are represented as mean±s.d. One-way analysis of variance isused to calculate the statistical significance of the results of allassays and P-values<0.05 are considered statistically significant.

Example 47 Study of the Effects of Compounds of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III) onIschemia/Reperfusion-induced Acute Kidney Injury

A study is performed to determine the effects of compounds of Formula(I), Formula (Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, onIschemia/Reperfusion-induced (I/R-induced) Acute Kidney Injury in CD-1(ICR) mice.

CD-1 (ICR) mice are purchased from Charles River Laboratory (Wilmington,Mass.). Mice are housed in a temperature- and humidity-controlledenvironment with a 12:12 hrs light-dark cycle and are allowed freelyaccess to standard rodent chow (TekLad, Madison, Wis.) and tap water.

Mice are subjected to a midline back incision, and both renal pediclesare clamped for 45 min with microaneurysm clamps (00396-01; Fine ScienceTools, Foster City, Calif.). After removal of the clamp, the kidneys areinspected for the restoration of blood flow. The animals are allowed torecover, and they are sacrificed 48 hrs after reperfusion. Mice aretreated with 100 mg/kg of a compound of Formula (I), Formula (Ia),Formula (Ib), Formula (II), or Formula (III), or a pharmaceuticallyacceptable salt thereof, by oral gavage once per day. CD-1 mice aredivided into four groups: (1) young mice with sham injury (n=4) (6-7weeks old); (2) young mice with I/R injury (n=8); (3) adult mice withsham injury (n=4) (20-24 weeks old); and (4) adult mice with I/R injury(n=11). An additional 27 adult mice (20-24 weeks old) are randomizedinto two groups: 13 mice received a compound of Formula (I), Formula(Ia), Formula (Ib), Formula (II), or Formula (III), or apharmaceutically acceptable salt thereof, and the other 14 mice receivedthe vehicle as a control.

The serum creatinine level is measured using the QuantiChrom CreatinineAssay Kit (DICT-500, BioAssay Systems, Hayward, Calif.). BUNmeasurements are recorded using the Infinity Urea (Nitrogen) LiquidStable Reagent (TR12421; ThermoTrace, Victoria, AU).

Evaluation of Renal Tissue

Kidneys are fixed in 4% paraformaldehyde, embedded in paraffin, andstained with hematoxylin and eosin (4 mm thick). Tubular injury isscored on a scale of 0-4 on the basis of the percentage of tubules withnecrosis, dilatation, or cell swelling: 0, less than 5%; 1, 5-25%; 2,25-50%; 3, 50-75%; and 4, over 75%. All high-power fields (×400) in thecortex and outer medulla are evaluated by a pathologist in a blindedmanner.

All values are expressed as mean±s.e. Statistical analysis is carriedout using GraphPad Prism 4.00 (San Diego, Calif.) with unpairedStudent's t testing for two sets of data and an analysis of variancewith a Bonferroni post-test for multiple groups. P<0.05 was consideredsignificant.

Example 48 Determination of the Effects of Compounds 1 and 17 on FoxO1Phosphorylation Levels

AML-12 cells were treated with different concentrations of Compound 1 orCompound 17 for 24 hours. Cells were then lysed in lysis buffer (50 mMTris, 150 mM KCl, EDTA 1 mM, NP40 1%) containing protease andphosphatase inhibitors, and analyzed by SDS-PAGE/western blot. Blockingand antibody incubations were done in 5% milk. Each protein present wasdetected with its specific antibody. Tubulin antibody was obtained fromSigma Inc, FoxO1 and phopho-FoxO1 (Ser256) antibodies were obtained fromCell Signaling. Antibody detection reactions were developed by enhancedchemiluminescence (Advansta, Calif., USA) using x-ray films.

FoxO1 phosphorylation at Ser256 results in its nuclear export and ininhibition of its transcription factor activity. A decrease in FoxO1phosphorylation at Ser256 with increasing dose of Compound 1 and 17 wasobserved (FIG. 6), indicating an increase in nuclear-translocated FoxO1and therefore increased FoxO1 transcriptional activity.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this disclosure belongs. In the specification, thesingular forms also include the plural unless the context clearlydictates otherwise. Although methods and materials similar or equivalentto those described herein can be used in the practice of testing thepresent disclosure, suitable methods and materials are described below.All publications, patent applications, patents, and other referencesmentioned herein are hereby expressly incorporated by reference. Thereferences cited herein are not admitted to be prior art of the claimeddisclosure. In the case of conflict, the present specification,including definitions, will control. In addition, the materials,methods, and examples are illustrative only and are not intended to belimiting.

Example 49 Synthesis of Exemplified Compounds

See, Hirose M, et al.,. “Design and synthesis of novel DFG-outRAF/vascular endothelial growth factor receptor 2 (VEGFR2) inhibitors:3. Evaluation of 5-amino-linked thiazolo[5,4-d]pyrimidine andthiazolo[5,4-b]pyridine derivatives.” Bioorg. Med. Chem. 2012, 15;20(18):5600-15.

See, Clift M D, Silverman R B., “Synthesis and evaluation of novelaromatic substrates and competitive inhibitors of GABAaminotransferase,” Bioorg. Med. Chem. Lett. 2008, 15; 18(10):3122-5.

A. M. El-Reedy, A. O. Ayyad and A. S. Ali, “Azolopyrimidines andpyrimidoquinazolines from 4-chloropyrimidines,” J. Het. Chem. 1989, 26,313-16.

See, Iwahashi M, et al., “Design and synthesis of new prostaglandin D₂receptor antagonists,” Bioorg. Med. Chem. 2011, 19(18):5361-71.

See, U.S. 2008/004,302(A1); and U.S. Pat. No. 8,716,470 (B2).

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments and methods described herein. Such equivalents are intendedto be encompassed by the scope of the present disclosure.

The invention claimed is:
 1. A compound of Formula (I):

or a pharmaceutically acceptable salt or tautomer thereof, wherein: X isO, OH, or Cl; L is —(CH₂)_(m)CH₂CH₂—, —(CH₂)_(m)Y(CH₂)_(p)—,—(CH₂)_(m)C(O)(CH₂)_(p)—, —(CH₂)_(m)C(O)O(CH₂)_(p)—,—(CH₂)_(m)C(O)NR²(CH₂)_(p)—, or —(CH₂)_(m)NR²C(O)(CH₂)_(p); Y is O orS(O)_(q); R¹ is C₆-C₁₀ aryl or heteroaryl, wherein the aryl andheteroaryl are substituted with R^(a) and R^(b), and optionallysubstituted with one or more R^(e); R² is H or C₁-C₆ alkyl; one of R^(a)and R^(b) is hydrogen and the other is —(CH₂)_(r)CO₂R^(x),—OCH₂CO₂R^(x), —(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)dihydrotetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r)isoxazol-3-ol, —(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH,—(CH₂)_(r)C(O)NHCN, or —(CH₂)_(r)C(O)NHS(O)₂alkyl; R^(c) is C₁-C₆haloalkyl, halogen, —CN, —OR^(x),or —CO₂R^(x); R^(d) is methyl,optionally substituted 5- to 10-membered aryl, optionally substituted 5-or 6-membered heteroaryl, or optionally substituted 5- or 6-memberedcarbocycle; each R^(x) is independently at each occurrence hydrogen orC₁-C₆ alkyl; each R^(e) is independently C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, halogen, —OR^(y), C₁-C₆ haloalkyl, —NHR^(z), —OH, or —CN;R^(f) is H or absent; each R^(y) and R^(z) is independently hydrogen,C₁-C₆ alkyl, or C₁-C₆ haloalkyl; each m and p independently is 0, 1 or2, wherein m+p<3; q is 0, 1, or 2; r is 0 or 1; and the dotted line isan optional double bond; with the proviso that R^(c) is not —CN when Xis O, L is —SCH₂— and R^(d) is optionally substituted phenyl; and thatR^(c) is not —CN when X is O, L is —SCH₂— and R^(d) is 2-furyl.
 2. Thecompound of claim 1, having Formula (Ia):

or a pharmaceutically acceptable salt, or tautomer thereof.
 3. Thecompound of claim 1, having Formula (Ib):

or a pharmaceutically acceptable salt thereof, wherein: one of R^(a) andR^(b) is hydrogen and the other is CO₂R^(x), CH₂CO₂R^(x), tetrazole, oroxadiazolone; R^(c) is halogen, —CN, or —OR^(x); R^(d) is methyl,optionally substituted 5- to 10-membered aryl, optionally substituted 5-or 6-membered heteroaryl, or optionally substituted 5- or 6-memberedcarbocycle; and R^(x) is hydrogen or C₁-C₆ alkyl; each R^(e) isindependently C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, halogen,—OR^(y), C₁-C₆ haloalkyl, —NHR^(z), —OH, or —CN; each R^(y) and R^(z) isindependently hydrogen, C₁-C₆ alkyl, or C₁-C₆ haloalkyl; and n is 0, 1,2, or 3; with the proviso that R^(c) is not —CN when R^(d) is optionallysubstituted phenyl and that R^(c) is not —CN when R^(d) is 2-furyl. 4.The compound of claim 1, having Formula (II):

or a pharmaceutically acceptable salt thereof, wherein R^(c) is halogen,—CN, or —OR^(x), and R^(d) is methyl, optionally substituted 5- to10-membered aryl, optionally substituted 5- or 6-membered heteroaryl, oroptionally substituted 5- or 6-membered carbocycle, and R^(x) ishydrogen or C₁-C₆ alkyl.
 5. The compound of claim 1, wherein R^(d) ismethyl, cyclohexyl, pyridinyl, thiazolyl, phenyl, or thienyl.
 6. Thecompound of claim 1, wherein R^(d) is methyl, cyclohexyl, pyridinyl,thiazolyl, thienyl, or optionally substituted phenyl.
 7. The compound ofclaim 1, wherein R^(a) is hydrogen, CH₂CO₂H, tetrazole, or oxadiazolone(1,2,4-oxadiazol-5(4H)-one).
 8. The compound of claim 1, wherein R^(b)is hydrogen, CH₂CO₂H, tetrazole, or oxadiazolone(1,2,4-oxadiazol-5(4H)-one).
 9. The compound of claim 1, having Formula(III)

or a pharmaceutically acceptable salt thereof, wherein: R^(a) and R^(b)is hydrogen and the other is —(CH₂)_(r)CO₂R^(x), —OCH₂CO₂R^(x),—(CH₂)_(r)tetrazole, —(CH₂)_(r)oxadiazolone,—(CH₂)_(r)dihydrotetrazolone, —(CH₂)_(r)thiadiazolol, —(CH₂)_(r)isoxazol-3-ol, —(CH₂)_(r)P(O)(OH)OR^(x), —(CH₂)_(r)S(O)₂OH,—(CH₂)_(r)C(O)NHCN, or —(CH₂)_(r)C(O)NHS(O)₂alkyl; R^(c) is C₁-C₆ alkyl,C₁-C₆ haloalkyl, halogen, —CN, —OR^(x), or —CO₂R^(x); R^(d) is methyl,optionally substituted 5- to 10-membered aryl, optionally substituted 5-or 6-membered heteroaryl, or optionally substituted 5- or 6-memberedcarbocycle; each R^(x) is independently at each occurrence hydrogen orC₁-C₆ alkyl; each R^(e) is independently C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, halogen, —OR^(y), C₁-C₆ haloalkyl, —NHR^(z), —OH, or —CN;each R^(y) and R^(z) is independently hydrogen, C₁-C₆ alkyl, or C₁-C₆haloalkyl; n is 0, 1, 2, or
 3. 10. A compound selected from the groupconsisting of:

or a pharmaceutically acceptable salt thereof.
 11. A pharmaceuticalcomposition comprising a compound of claim 1, or a pharmaceuticallyacceptable salt thereof, and at least one of a pharmaceuticallyacceptable carrier, diluent, or excipient.
 12. The pharmaceuticalcomposition according to claim 11, further comprising one or moreadditional therapeutic agents.
 13. The compound of claim 1, where R^(c)is —CN or halogen.